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1
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Wu Y, Liu C, Yao H, He S, Zhao L, Zeng X. Investigation of the factors influencing the selectivity and response times of glyoxal-selective probes. Org Biomol Chem 2025; 23:5139-5147. [PMID: 40298951 DOI: 10.1039/d5ob00366k] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 04/30/2025]
Abstract
In this work, four o-phenylenediamine-conjugated fluorescent probes 1-4 with different substituents on the amino group have been designed and prepared to investigate the factors influencing the selectivity and response times of the probes toward glyoxal (GO). The introduced substituted groups, including n-propyl, iso-propyl, tert-butyl, and phenyl, 1-4 displayed sufficient selectivity toward GO. Due to the fact that GO normally exists in a hydrated state in aqueous media, the reaction rates between 1-4 and GO exhibited significant deviation from the steric hindrance effects of most organic reactions. Compounds 3 and 4 with bulky substituted groups presented faster reaction rates with GO than those of compounds 1-2. To further elucidate the properties of these compounds, we selected compound 3 as the representative compound, because of its best selectivity and fast response time, for detailed investigation of the recognition behaviors both in aqueous solution and in living cells. Probe 3 exhibited high selectivity and high sensitivity toward GO with a large Stokes shift (>100 nm), making it highly suitable for in vivo imaging research.
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Affiliation(s)
- Yuanyuan Wu
- Tianjin Key Laboratory for Photoelectric Materials and Devices, School of Materials Science & Engineering, Tianjin University of Technology, Tianjin 300384, China.
- Key Laboratory of Display Materials and Photoelectric Devices, Ministry of Education, School of Materials Science & Engineering, Tianjin University of Technology, Tianjin, 300384, China
| | - Chang Liu
- Tianjin Key Laboratory for Photoelectric Materials and Devices, School of Materials Science & Engineering, Tianjin University of Technology, Tianjin 300384, China.
- Key Laboratory of Display Materials and Photoelectric Devices, Ministry of Education, School of Materials Science & Engineering, Tianjin University of Technology, Tianjin, 300384, China
| | - Huirong Yao
- Tianjin Key Laboratory for Photoelectric Materials and Devices, School of Materials Science & Engineering, Tianjin University of Technology, Tianjin 300384, China.
- Key Laboratory of Display Materials and Photoelectric Devices, Ministry of Education, School of Materials Science & Engineering, Tianjin University of Technology, Tianjin, 300384, China
| | - Song He
- Tianjin Key Laboratory for Photoelectric Materials and Devices, School of Materials Science & Engineering, Tianjin University of Technology, Tianjin 300384, China.
- Key Laboratory of Display Materials and Photoelectric Devices, Ministry of Education, School of Materials Science & Engineering, Tianjin University of Technology, Tianjin, 300384, China
| | - Liancheng Zhao
- School of Materials Science and Engineering, Institute of Information Functional Materials& Devices, Harbin Institute of Technology, Harbin, 150001, China
| | - Xianshun Zeng
- Tianjin Key Laboratory for Photoelectric Materials and Devices, School of Materials Science & Engineering, Tianjin University of Technology, Tianjin 300384, China.
- Key Laboratory of Display Materials and Photoelectric Devices, Ministry of Education, School of Materials Science & Engineering, Tianjin University of Technology, Tianjin, 300384, China
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2
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Kopera M, Adamkiewicz M, Pieniazek A. Effect of Glyoxal on Plasma Membrane and Cytosolic Proteins of Erythrocytes. Int J Mol Sci 2025; 26:4328. [PMID: 40362565 PMCID: PMC12072774 DOI: 10.3390/ijms26094328] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/25/2025] [Revised: 04/28/2025] [Accepted: 04/29/2025] [Indexed: 05/15/2025] Open
Abstract
Glyoxal (GO) is a reactive dicarbonyl derived endogenously from sugars and other metabolic reactions within cells. Numerous exogenous sources of this compound include tobacco smoking, air pollution, and food processing. GO is toxic to cells mainly due to its high levels and reactivity towards proteins, lipids, and nucleic acids. We speculate that glyoxal could be involved in erythrocyte protein damage and lead to cell dysfunction. The osmotic fragility and level of amino and carbonyl groups of membrane proteins of erythrocytes incubated for 24 h with GO were identified. The amount of thiol, amino, and carbonyl groups was also measured in hemolysate proteins after erythrocyte treatment with GO. In hemolysate, the level of glutathione, non-enzymatic antioxidant capacity (NEAC), TBARS, and activity of antioxidant enzymes was also determined. The study's results indicated that GO increases erythrocyte osmotic sensitivity, alters the levels of glutathione and free functional groups in hemolysate proteins, and modifies the activity of antioxidant enzymes. Our findings indicate that GO is a highly toxic compound to human erythrocytes. Glyoxal at concentrations above 5 mM can cause functional changes in erythrocyte proteins and disrupt the oxidoreductive balance in cells.
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Affiliation(s)
- Michal Kopera
- Department of Oncobiology and Epigenetics, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143 Str., 90-236 Lodz, Poland; (M.K.); (M.A.)
- Doctoral School of Exact and Natural Sciences, University of Lodz, 21/23 Jana Matejki Str., 90-236 Lodz, Poland
| | - Malgorzata Adamkiewicz
- Department of Oncobiology and Epigenetics, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143 Str., 90-236 Lodz, Poland; (M.K.); (M.A.)
| | - Anna Pieniazek
- Department of Oncobiology and Epigenetics, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143 Str., 90-236 Lodz, Poland; (M.K.); (M.A.)
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3
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Yang Y, Hu X. The Nearly Complete Genome of Grifola frondosa and Light-Induced Genes Screened Based on Transcriptomics Promote the Production of Triterpenoid Compounds. J Fungi (Basel) 2025; 11:322. [PMID: 40278142 PMCID: PMC12028448 DOI: 10.3390/jof11040322] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/02/2025] [Revised: 04/10/2025] [Accepted: 04/16/2025] [Indexed: 04/26/2025] Open
Abstract
Grifola frondosa, commonly known as turkey tail, is a valuable fungus with medicinal and culinary uses, rich in bioactive compounds like triterpenoid polysaccharides that contribute to health benefits. Here, we constructed a nearly complete genome of G. frondosa CH1 using Illumina, PacBio HiFi, and Hi-C sequencing technologies, resulting in a 35.74 Mb genome with 12,526 protein-coding genes. The genome spans 12 chromosomes, all with intact telomeric structures and no gaps. The BUSCO completeness scores of 95.1% and 99.1% for the genome and genes, respectively, indicate high assembly quality and high completeness of gene prediction. Phylogenetic analysis showed a close relationship between G. frondosa CH1 and Trametes cinnabarina. Transcriptomic analysis under varying light conditions showed changes in the expression of genes, especially those related to terpenoid synthesis, with several CAZymes and CYP450 genes also exhibiting light-induced variations. Ten triterpenoid secondary metabolite gene clusters were identified, three of which were light-sensitive, indicating that light exposure regulates triterpenoid metabolism. This study provides valuable data supporting the high-quality genome of G. frondosa and offers new insights into the light-induced regulation of its metabolism.
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Affiliation(s)
- Yang Yang
- Institute for Medicinal Plants, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China;
- Innovation Academy of International Traditional Chinese Medicinal Materials, Huazhong Agricultural University, Wuhan 430070, China
| | - Xuebo Hu
- Institute for Medicinal Plants, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China;
- Innovation Academy of International Traditional Chinese Medicinal Materials, Huazhong Agricultural University, Wuhan 430070, China
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Qiao X, Zhang L, Hoffman EA, Mastin GE, Hileman CO, Kallianpur AR, Wang M, Ellis RJ, Koletar SL, Palella FJ, Tassiopoulos KK, Landay AL, Kapahi P, Galligan JJ, Kalayjian RC. Glycation metabolites predict incident age-related comorbidities and mortality in older people with HIV. GeroScience 2025:10.1007/s11357-025-01652-3. [PMID: 40240751 DOI: 10.1007/s11357-025-01652-3] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/09/2025] [Accepted: 04/02/2025] [Indexed: 04/18/2025] Open
Abstract
Glycation is a class of modifications arising from non-enzymatic reactions of reducing sugars with proteins, lipids, and/or DNA, generating advanced glycation end-products (AGEs). AGEs are linked to many age-related comorbidities. In response to HIV-1 infection, activated T-cells and macrophages shift their predominate metabolism from oxidative phosphorylation to glycolysis. Increased glycolytic flux enhances AGE formation, which may increase age-related comorbidities. In this prospective, multicenter cohort study of antiretroviral therapy treated people with HIV, we explored predictive associations by baseline plasma AGE concentrations and their corresponding detoxification metabolites, with incident comorbidities and mortality. AGEs included dicarbonyl sugars: 3-deoxyglucosone, glyoxal, and methylglyoxal. Methylglyoxal-derived metabolites included carboxyethyl-arginine, carboxyethyl-lysine, and methylglyoxal hydroimidazolone-1. Detoxification metabolites included reduced and oxidized glutathione, and the glyoxalase cycle products lactoyl-glutathione and lactoyl-Lysine modified proteins. Plasma was collected at study entry, in the fasting state, and assayed by liquid chromatography-mass spectroscopy. Incident clinical outcomes included diabetes, chronic kidney disease, hypertension, neurocognitive impairment, peripheral neuropathy, frailty, fractures, recurrent falls, and all-cause mortality. Among 376 participants, higher baseline plasma concentrations of methylglyoxal derived AGEs predicted increased risks of diabetes, chronic kidney disease, and recurrent falls, while higher 3-deoxyglucosone predicted an increased risk of peripheral neuropathy. By contrast, higher baseline concentrations of reduced or oxidized glutathione, lactoyl-glutathione, and/or lactoyl-Lysine modified proteins predicted lower risks of diabetes, neurocognitive impairment, frailty, fractures, recurrent falls, and all-cause mortality. These findings support growing experimental evidence of the potential to mitigate age-related declines by interventions that reduce glycation or increase glutathione.
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Affiliation(s)
- Xi Qiao
- Department of Population and Quantitative Health Sciences, Case Western Reserve Univ, Cleveland, OH, USA
| | - Liangliang Zhang
- Department of Population and Quantitative Health Sciences, Case Western Reserve Univ, Cleveland, OH, USA
- Case Comprehensive Cancer Center, Cleveland, OH, USA
| | - Emely A Hoffman
- Department of Pharmacology and Toxicology, University of Arizona, Tucson, AZ, USA
| | - Grace E Mastin
- Department of Pharmacology and Toxicology, University of Arizona, Tucson, AZ, USA
| | - Corrilynn O Hileman
- MetroHealth Medical Center, Cleveland, OH, USA
- Case Western Reserve University School of Medicine, Cleveland, OH, USA
| | | | - Ming Wang
- Department of Population and Quantitative Health Sciences, Case Western Reserve Univ, Cleveland, OH, USA
| | | | | | - Frank J Palella
- Northwestern University Feinberg School of Medicine, Chicago, IL, USA
| | | | - Alan L Landay
- University of Texas Medical Branch, Galveston, TX, USA
| | - Pankaj Kapahi
- The Buck Institute for Research on Aging, Novato, CA, USA
| | - James J Galligan
- Department of Pharmacology and Toxicology, University of Arizona, Tucson, AZ, USA.
| | - Robert C Kalayjian
- MetroHealth Medical Center, Cleveland, OH, USA.
- Case Western Reserve University School of Medicine, Cleveland, OH, USA.
- Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, OH, USA.
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5
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Kashihara T, Yasaki M, Okuyama Y, Murayama A, Morita A, Nakahara T. YAP activation in Müller cells alleviates oxidative stress in the rat retina after intravitreal injection with methylglyoxal. J Pharmacol Sci 2025; 157:219-228. [PMID: 40058941 DOI: 10.1016/j.jphs.2025.02.005] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/17/2024] [Revised: 01/27/2025] [Accepted: 02/22/2025] [Indexed: 05/13/2025] Open
Abstract
Methylglyoxal (MGO), a highly reactive dicarbonyl compound produced via the glycolytic pathway, plays a key role in the pathogenesis of various diabetic complications, such as diabetic retinopathy. Müller cells provide neurotrophic support and maintain retinal homeostasis, including the redox balance. This dysfunction leads to retinal disease. Yes-associated protein (YAP), a major downstream effector of the Hippo pathway, plays a crucial role in regulating cell survival. In this study, we investigated the roles of Müller cell YAP during MGO-induced retinal injury using normal rats intravitreally injected with MGO and a rat Müller cell line (rMC-1). Immunohistochemistry revealed that MGO injection increased the glial fibrillary acidic protein immunoreactivity in Müller cells. The alignment of Müller cell nuclei was disrupted in MGO-treated retinas. YAP increased and activated in Müller cells two days after MGO injection. This increase in YAP levels was independent of the Hippo pathway and partially attributed to the upregulation of YAP mRNA levels. YAP inhibition by verteporfin exacerbated MGO-induced cell damage and decreased Bcl-xL levels in rMC-1 cells. Intravitreal verteporfin injection also enhanced MGO-induced retinal oxidative stress. Overall, our findings suggest that YAP activation in Müller cells alleviates oxidative stress in the retina following MGO-induced retinal injury.
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Affiliation(s)
- Toshihide Kashihara
- Department of Molecular Pharmacology, Kitasato University School of Pharmaceutical Sciences, Tokyo, 108-8641, Japan.
| | - Mayuko Yasaki
- Department of Molecular Pharmacology, Kitasato University School of Pharmaceutical Sciences, Tokyo, 108-8641, Japan
| | - Yumi Okuyama
- Department of Molecular Pharmacology, Kitasato University School of Pharmaceutical Sciences, Tokyo, 108-8641, Japan
| | - Aki Murayama
- Department of Molecular Pharmacology, Kitasato University School of Pharmaceutical Sciences, Tokyo, 108-8641, Japan
| | - Akane Morita
- Department of Molecular Pharmacology, Kitasato University School of Pharmaceutical Sciences, Tokyo, 108-8641, Japan
| | - Tsutomu Nakahara
- Department of Molecular Pharmacology, Kitasato University School of Pharmaceutical Sciences, Tokyo, 108-8641, Japan.
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Vangrieken P, Scheijen JLJM, Schiffers PMH, van de Waarenburg MPH, Foulquier S, Schalkwijk CCG. Modelling the effects of elevated methylglyoxal levels on vascular and metabolic complications. Sci Rep 2025; 15:6025. [PMID: 39972072 PMCID: PMC11839914 DOI: 10.1038/s41598-025-90661-5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/06/2024] [Accepted: 02/14/2025] [Indexed: 02/21/2025] Open
Abstract
Methylglyoxal (MGO), a glycolysis by-product and precursor to advanced glycation endproducts (AGEs), is associated with glucose intolerance, type 2 diabetes, and vascular dysfunction. This study examined the long-term effects of elevated MGO on blood pressure, insulin sensitivity, and vascular function in healthy mice. Male C57Bl/6J mice were assigned to control (n = 16) or MGO-treated groups (50 mM in drinking water for 13 weeks, n = 16). Measurements included body weight, fasting plasma glucose, water consumption, blood pressure, and analysis of plasma/tissue for MGO, AGEs, glyoxalase activity, and inflammation markers. Endothelial function was assessed using wire myography, and the response of human placental arteries to MGO-modified insulin was evaluated. MGO treatment significantly increased plasma MGO (123.3%, p < 0.001), AGEs MG-H1 (208.6%, p < 0.001) and CEL (64.3%, p < 0.001), and AGEs in the heart, kidney, and liver, along with body weight (+ 6.4%, p = 0.032) and blood pressure (systolic + 5.0%, p = 0.046; diastolic + 6.5%, p = 0.043). Glucose sensitivity and endothelial function remained unaffected. CRP levels rose, and MGO-modified insulin enhanced vascular contraction. In conclusion, chronic MGO exposure increased plasma MGO to diabetic-like levels, raised body weight and blood pressure, and did not alter glucose sensitivity or endothelial function. Modification of insulin by MGO may contribute to MGO-related changes in blood pressure.
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Affiliation(s)
- Philippe Vangrieken
- CARIM, Cardiovascular Research Institute Maastricht, Department of Internal Medicine, School for Cardiovascular Diseases (CARIM), Maastricht University Medical Center+, Universiteitssingel 50, 6229 ER, Maastricht, The Netherlands.
- CARIM, Cardiovascular Research Institute Maastricht, Department of Pharmacology and Toxicology, CARIM, Maastricht University Medical Center+, Maastricht, The Netherlands.
| | - Jean L J M Scheijen
- CARIM, Cardiovascular Research Institute Maastricht, Department of Internal Medicine, School for Cardiovascular Diseases (CARIM), Maastricht University Medical Center+, Universiteitssingel 50, 6229 ER, Maastricht, The Netherlands
| | - Paul M H Schiffers
- CARIM, Cardiovascular Research Institute Maastricht, Department of Pharmacology and Toxicology, CARIM, Maastricht University Medical Center+, Maastricht, The Netherlands
| | - Marjo P H van de Waarenburg
- CARIM, Cardiovascular Research Institute Maastricht, Department of Internal Medicine, School for Cardiovascular Diseases (CARIM), Maastricht University Medical Center+, Universiteitssingel 50, 6229 ER, Maastricht, The Netherlands
| | - Sebastien Foulquier
- CARIM, Cardiovascular Research Institute Maastricht, Department of Pharmacology and Toxicology, CARIM, Maastricht University Medical Center+, Maastricht, The Netherlands
- MHeNs, Mental Health and Neuroscience Research Institute, Maastricht University Medical Center+, Maastricht, The Netherlands
| | - Casper C G Schalkwijk
- CARIM, Cardiovascular Research Institute Maastricht, Department of Internal Medicine, School for Cardiovascular Diseases (CARIM), Maastricht University Medical Center+, Universiteitssingel 50, 6229 ER, Maastricht, The Netherlands
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7
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Wei X, Sun Z, Wang N, Deng Z, Li W, Ying T, Wu M, Liu Y, He G. Immunometabolic profiling related with gestational diabetes mellitus: a nested case-control study of CD4 + T cell phenotypes and glycemic traits. Acta Diabetol 2025; 62:77-85. [PMID: 39147954 DOI: 10.1007/s00592-024-02338-6] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/07/2024] [Accepted: 07/06/2024] [Indexed: 08/17/2024]
Abstract
AIMS To investigate immunometabolic associations of CD4+ T cell phenotypes with gestational diabetes mellitus (GDM). METHODS A nested case-control study was conducted comprising 53 pairs of GDM patients and matched controls within a prospective cohort. Metabolomic signatures related to both CD4+ T cell phenotypes and glycemic traits among pregnant women were investigated by weighted gene co-expression network analysis (WGCNA). Multivariable-adjusted generalized linear models were used to explore the associations of CD4+ T cell phenotypes and selected metabolites with GDM. Mediation analysis was conducted to evaluate the mediating effect of selected metabolites on the relationship between CD4+ T cell phenotypes and glycemic traits. RESULTS Higher levels of Treg cells (OR per SD increment (95%CI): 0.57 (0.34, 0.95), p = 0.031) and increased expression of Foxp3 (OR per SD increment (95%CI): 0.59 (0.35, 0.97), p = 0.039) and GATA3 (OR per SD increment (95%CI): 0.42 (0.25, 0.72), p = 0.002) were correlated with a decreased risk of GDM. Plasma pyruvaldehyde, S-adenosylhomocysteine (SAH), bergapten, and 9-fluorenone mediated the association between Tregs and fasting plasma glucose (FPG), with mediation proportions of 46.9%, 39.6%, 52.4%, and 56.9%, respectively. CONCLUSIONS Treg cells and Foxp3 expressions were inversely associated with GDM risk, with potential metabolic mechanisms involving metabolites such as pyruvaldehyde and SAH.
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Affiliation(s)
- Xiaohui Wei
- School of Public Health, Key Laboratory of Public Health Safety, Ministry of Education, Fudan University, No.130, Dong'an Road, Xuhui District, Shanghai, 200032, China
| | - Zhuo Sun
- School of Public Health, Key Laboratory of Public Health Safety, Ministry of Education, Fudan University, No.130, Dong'an Road, Xuhui District, Shanghai, 200032, China
| | - Na Wang
- Nursing Department, Obstetrics and Gynaecology Hospital of Fudan University, Shanghai, 200011, China
| | - Zequn Deng
- School of Public Health, Key Laboratory of Public Health Safety, Ministry of Education, Fudan University, No.130, Dong'an Road, Xuhui District, Shanghai, 200032, China
| | - Wenyun Li
- School of Public Health, Key Laboratory of Public Health Safety, Ministry of Education, Fudan University, No.130, Dong'an Road, Xuhui District, Shanghai, 200032, China
| | - Tao Ying
- School of Public Health, Key Laboratory of Public Health Safety, Ministry of Education, Fudan University, No.130, Dong'an Road, Xuhui District, Shanghai, 200032, China
| | - Min Wu
- School of Public Health, Key Laboratory of Public Health Safety, Ministry of Education, Fudan University, No.130, Dong'an Road, Xuhui District, Shanghai, 200032, China
| | - Yuwei Liu
- School of Public Health, Key Laboratory of Public Health Safety, Ministry of Education, Fudan University, No.130, Dong'an Road, Xuhui District, Shanghai, 200032, China.
| | - Gengsheng He
- School of Public Health, Key Laboratory of Public Health Safety, Ministry of Education, Fudan University, No.130, Dong'an Road, Xuhui District, Shanghai, 200032, China.
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Jiang W, Ma X, Li B, Jiang T, Jiang H, Chen W, Gao J, Mao Y, Sun X, Ye Z, Zhao S, Huang S, Chen Y. Role of the PGAM5-CypD mitochondrial pathway in methylglyoxal-induced bone loss in diabetic osteoporosis. Bone 2024; 190:117322. [PMID: 39510433 DOI: 10.1016/j.bone.2024.117322] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/27/2024] [Revised: 10/24/2024] [Accepted: 11/04/2024] [Indexed: 11/15/2024]
Abstract
Diabetic osteoporosis (DOP) is a skeletal complication with a high rate of disability. It results in a great burden to the patient's family and society. Methylglyoxal (MG) is a toxic by-product of the glycolytic process that occurs during diabetic conditions. It causes osteoblastic injury and con-tributes to the initiation and development of DOP. Disruption of mitochondrial homeostasis has been implicated as a cause of dysregulated osteo-blastogenesis, an essential step in bone formation. It is unclear whether mitochondrial dysfunction is involved in MG-induced osteoblast dysfunction. In this study, we showed that mitochondrial dysfunction contributes to MG-induced MC3T3-E1 cell apoptosis and impaired differentiation. A significant reduction of mitochondrial membrane potential (MMP) and ATP production occurred in MG-induced osteoblasts as well as increasing mitochondrial reactive oxygen species (mtROS) and intracellular Ca2+. Classical antioxidant N-Acetylcysteine (NAC) significantly attenuated mitochondrial dysfunction as well as osteoblast apoptosis and osteogenic differentiation damage induced by MG. More importantly, we found that activating phosphoglycerate mutase family member 5 (PGAM5) and cyclophilin D (CypD), which contributes to mitochondrial homeostasis, is involved in MG-induced osteoblast injury. Both PGAM5 and CypD knockdown effectively reversed osteoblast viability and function, whereas PGAM5 or CypD overexpression aggravated osteoblast injury caused by MG. Moreover, the result of co-transfection revealed that PGAM5 is an upstream signaling molecule of CypD. By constructing type I diabetes mouse models, we further found that the expression of PGAM5 and CypD were both increased in the femur along with a reduction of ATP and increased TUNEL-positive cells. These results, for the first time, suggest that MG-induced mitochondrial dysfunction induces osteoblast injury through the PGAM5-CypD pathway. This study provides insight into the prevention and treatment of DOP. LAY SUMMARY: This study highlights the role of mitochondria in regulating osteoblast viability and function under conditions of diabetic osteoporosis (DOP). We found that the PGAM5-CypD mitochondrial pathway is activated following glycolytic by-product methylglyoxal (MG) treatment, which contributes to mitochondrial dysfunction and osteogenic dysfunction. This mechanism implicates mitochondria as a potential therapeutic target for osteoporosis.
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Affiliation(s)
- Wanying Jiang
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China
| | - Xinyi Ma
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China
| | - Bin Li
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China
| | - Tianle Jiang
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China
| | - Haopu Jiang
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China
| | - Wenxia Chen
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China
| | - Jia Gao
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China
| | - Yixin Mao
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China; Department of Prosthodontics, School and Hospital of Stomatology, Wenzhou Medical University, China
| | - Xiaoyu Sun
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China; Department of Periodontics, School and Hospital of Stomatology, Wenzhou Medical University, China
| | - Zhou Ye
- Applied Oral Sciences and Community Dental Care, Faculty of Dentistry, University of Hong Kong, Hong Kong
| | - Shufan Zhao
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China; Department of Oral Maxillofacial Surgery, School and Hospital of Stomatology, Wenzhou Medical University, China.
| | - Shengbin Huang
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China; Department of Prosthodontics, School and Hospital of Stomatology, Wenzhou Medical University, China.
| | - Yang Chen
- Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, China; Department of Prosthodontics, School and Hospital of Stomatology, Wenzhou Medical University, China.
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9
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Thomas LA, Hopkinson RJ. The biochemistry of the carcinogenic alcohol metabolite acetaldehyde. DNA Repair (Amst) 2024; 144:103782. [PMID: 39566398 DOI: 10.1016/j.dnarep.2024.103782] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/22/2024] [Revised: 10/14/2024] [Accepted: 10/30/2024] [Indexed: 11/22/2024]
Abstract
Acetaldehyde (AcH) is the first metabolite of ethanol and is proposed to be responsible for the genotoxic effects of alcohol consumption. As an electrophilic aldehyde, AcH can form multiple adducts with DNA and other biomolecules, leading to function-altering and potentially toxic and carcinogenic effects. In this review, we describe sources of AcH in humans, including AcH biosynthesis mechanisms, and outline the structures, properties and functions of AcH-derived adducts with biomolecules. We also describe human AcH detoxification mechanisms and discuss ongoing challenges in the field.
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Affiliation(s)
- Liam A Thomas
- Institute of Structural and Chemical Biology, School of Chemistry, University of Leicester, Leicester LE1 7RH, UK
| | - Richard J Hopkinson
- Institute of Structural and Chemical Biology, School of Chemistry, University of Leicester, Leicester LE1 7RH, UK.
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10
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Zhong Y, Yang C, Lu Y, Lv L. Theanine Capture of Reactive Carbonyl Species in Humans after Consuming Theanine Capsules or Green Tea. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 2024; 72:13240-13249. [PMID: 38825967 DOI: 10.1021/acs.jafc.4c02481] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/04/2024]
Abstract
Acrolein (ACR), methylglyoxal (MGO), and glyoxal (GO) are a class of reactive carbonyl species (RCS), which play a crucial role in the pathogenesis of chronic and age-related diseases. Here, we explored a new RCS inhibitor (theanine, THE) and investigated its capture capacity on RCS in vivo by human experiments. After proving that theanine could efficiently capture ACR instead of MGO/GO by forming adducts under simulated physiological conditions, we further detected the ACR/MGO/GO adducts of theanine in the human urine samples after consumption of theanine capsules (200 and 400 mg) or green tea (4 cups, containing 200 mg of theanine) by using ultraperformance liquid chromatography-time-of-flight-high-resolution mass spectrometry. Quantitative assays revealed that THE-ACR, THE-2ACR-1, THE-MGO, and THE-GO were formed in a dose-dependent manner in the theanine capsule groups; the maximum value of the adducts of theanine was also tested. Furthermore, besides the RCS adducts of theanine, the RCS adducts of catechins could also be detected in the drinking tea group. Whereas, metabolite profile analysis showed that theanine could better capture RCS produced in the renal metabolic pathway than catechins. Our findings indicated that theanine could reduce RCS in the body in two ways: as a pure component or contained in tea leaves.
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Affiliation(s)
- Yuqing Zhong
- Department of Food Science and Technology, School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, 2# Xuelin Road, Nanjing 210023, P. R. China
| | - Chen Yang
- Department of Food Science and Technology, School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, 2# Xuelin Road, Nanjing 210023, P. R. China
| | - Yongling Lu
- Department of Food Science and Technology, School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, 2# Xuelin Road, Nanjing 210023, P. R. China
| | - Lishuang Lv
- Department of Food Science and Technology, School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, 2# Xuelin Road, Nanjing 210023, P. R. China
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11
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Vizuete AFK, Fróes F, Seady M, Hansen F, Ligabue-Braun R, Gonçalves CA, Souza DO. A Mechanism of Action of Metformin in the Brain: Prevention of Methylglyoxal-Induced Glutamatergic Impairment in Acute Hippocampal Slices. Mol Neurobiol 2024; 61:3223-3239. [PMID: 37980327 DOI: 10.1007/s12035-023-03774-1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/24/2023] [Accepted: 11/05/2023] [Indexed: 11/20/2023]
Abstract
Metformin, a biguanide compound (N-1,1-dimethylbiguanide), is widely prescribed for diabetes mellitus type 2 (T2D) treatment. It also presents a plethora of properties, such as anti-oxidant, anti-inflammatory, anti-apoptosis, anti-tumorigenic, and anti-AGE formation activity. However, the precise mechanism of action of metformin in the central nervous system (CNS) needs to be clarified. Herein, we investigated the neuroprotective role of metformin in acute hippocampal slices exposed to methylglyoxal (MG), a highly reactive dicarbonyl compound and a key molecule in T2D developmental pathophysiology. Metformin protected acute hippocampal slices from MG-induced glutamatergic neurotoxicity and neuroinflammation by reducing IL-1β synthesis and secretion and RAGE protein expression. The drug also improved astrocyte function, particularly with regard to the glutamatergic system, increasing glutamate uptake. Moreover, we observed a direct effect of metformin on glutamate transporters, where the compound prevented glycation, by facilitating enzymatic phosphorylation close to Lys residues, suggesting a new neuroprotective role of metformin via PKC ζ in preventing dysfunction in glutamatergic system induced by MG.
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Affiliation(s)
- Adriana Fernanda K Vizuete
- Laboratory of Calcium-Binding Proteins in the CNS, Department of Biochemistry, Institute of Basic Health Sciences, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, RS, Brazil.
- Post Graduate Program in Biochemistry, Institute of Basic Health Sciences, UFRGS, Porto Alegre, RS, Brazil.
- Department of Biochemistry, Institute of Basic Health Sciences, UFRGS, Ramiro Barcelos, 2600-Anexo, Porto Alegre, RS, 90035-003, Brazil.
| | - Fernanda Fróes
- Laboratory of Calcium-Binding Proteins in the CNS, Department of Biochemistry, Institute of Basic Health Sciences, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, RS, Brazil
- Post Graduate Program in Biochemistry, Institute of Basic Health Sciences, UFRGS, Porto Alegre, RS, Brazil
| | - Marina Seady
- Laboratory of Calcium-Binding Proteins in the CNS, Department of Biochemistry, Institute of Basic Health Sciences, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, RS, Brazil
- Post Graduate Program in Biochemistry, Institute of Basic Health Sciences, UFRGS, Porto Alegre, RS, Brazil
| | - Fernanda Hansen
- Department of Nutrition, Health Sciences Center, Federal University of Santa Catarina, University Campus, Trindade, Florianópolis, Santa Catarina, 88040-900, Brazil
| | - Rodrigo Ligabue-Braun
- Department of Pharmacosciences, Federal University of Health Sciences of Porto Alegre (UFCSPA), Avenida Sarmento Leite 245, Porto Alegre, 90050-130, Brazil
| | - Carlos-Alberto Gonçalves
- Laboratory of Calcium-Binding Proteins in the CNS, Department of Biochemistry, Institute of Basic Health Sciences, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, RS, Brazil
- Post Graduate Program in Biochemistry, Institute of Basic Health Sciences, UFRGS, Porto Alegre, RS, Brazil
- Department of Biochemistry, Institute of Basic Health Sciences, UFRGS, Ramiro Barcelos, 2600-Anexo, Porto Alegre, RS, 90035-003, Brazil
| | - Diogo O Souza
- Post Graduate Program in Biochemistry, Institute of Basic Health Sciences, UFRGS, Porto Alegre, RS, Brazil
- Department of Biochemistry, Institute of Basic Health Sciences, UFRGS, Ramiro Barcelos, 2600-Anexo, Porto Alegre, RS, 90035-003, Brazil
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12
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Kotian NP, Prabhu A, Tender T, Raghu Chandrashekar H. Methylglyoxal Induced Modifications to Stabilize Therapeutic Proteins: A Review. Protein J 2024; 43:39-47. [PMID: 38017314 DOI: 10.1007/s10930-023-10166-w] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 10/10/2023] [Indexed: 11/30/2023]
Abstract
Therapeutic proteins are potent, fast-acting drugs that are highly effective in treating various conditions. Medicinal protein usage has increased in the past 10 years, and it will evolve further as we better understand disease molecular pathways. However, it is associated with high processing costs, limited stability, difficulty in being administered as an oral medication, and the inability of large proteins to penetrate tissue and reach their target locations. Many methods have been developed to overcome the problems with the stability and chaperone activity of therapeutic proteins, viz., the addition of external agents (changing the properties of the surrounding solvent by using stabilizing excipients, e.g., amino acids, sugars, polyols) and internal agents (chemical modifications that influence its structural properties, e.g., mutations, glycosylation). However, these methods must completely clear protein instability and chaperone issues. There is still much work to be done on finetuning chaperone proteins to increase their biological efficacy and stability. Methylglyoxal (MGO), a potent dicarbonyl compound, reacts with proteins and forms covalent cross-links. Much research on MGO scavengers has been conducted since they are known to alter protein structure, which may result in alterations in biological activity and stability. MGO is naturally produced within our body, however, its impact on chaperones and protein stability needs to be better understood and seems to vary based on concentration. This review highlights the efforts of several research groups on the effect of MGO on various proteins. It also addresses the impact of MGO on a client protein, α-crystallin, to understand the potential solutions to the protein's chaperone and stability problems.
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Affiliation(s)
- Nainika Prashant Kotian
- Department of Pharmaceutical Biotechnology, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal, 576104, India
| | - Anusha Prabhu
- Department of Pharmaceutical Biotechnology, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal, 576104, India
| | - Tenzin Tender
- Department of Pharmaceutical Biotechnology, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal, 576104, India
| | - Hariharapura Raghu Chandrashekar
- Department of Pharmaceutical Biotechnology, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal, 576104, India.
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13
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Shahnawaz Khan M, Ahmad Bhat S, Saud Albagmi M, Arshad M, Tarique M, Bano B. Antiglycation potential of metal ions and polyphenolic extract of chickpea on thiol-protease inhibitor: A management for diabetic complications. Saudi Pharm J 2024; 32:101916. [PMID: 38178850 PMCID: PMC10765112 DOI: 10.1016/j.jsps.2023.101916] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/22/2023] [Accepted: 12/11/2023] [Indexed: 01/06/2024] Open
Abstract
Glycation is the non-enzymatic adduct formation between reducing sugars or dicarbonyls with proteins and is a crucial molecular event under hyperglycaemic conditions of diabetes. The accumulation of advanced glycation end products (AGEs) due to glycation of proteins has been implicated in several diseases associated with ageing and diabetes. Thus, investigating the antiglycation potential of some trace metal ions (Manganese; Mn2+, and Zinc; Zn2+) and polyphenolic extract of chickpea seeds (PEC) on the methylglyoxal (MGO) induced glycation of a phytocystatin isolated from chickpea was taken up to find an inexpensive and non-toxic therapeutic means of medicating protein glycation and associated diabetic complications. The current study focused on the comparative analyses of these micronutrients and herbal extracts in inhibiting protein glycation and AGEs formation in a quest to develop nutraceuticals for managing diabetes. The effect of metals (Mn2+, Zn2+) and PEC on protein glycation was assessed by different techniques, i.e., glycation-specific AGE fluorescence and absorbance, thiol protease inhibitory activity assay, and conformational alterations by spectroscopic assays. This study revealed the significant anti-glycation potencies of Mn2+, Zn2+, and PEC against the MGO-induced glycation of CPC, which might pave the way for resolving pathological complications of diabetes by combining higher levels of efficacy, selectivity, and safety in humans. Moreover, characterization and identification of different AGEs formed during the glycation process in diabetics was done to apply the same for determining the onset of glycation at the early stage so that appropriate steps be taken to address the menace of diabetic complications.
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Affiliation(s)
- Mohd Shahnawaz Khan
- Department of Biochemistry, College of Science, King Saud University, Riyadh, 11451, Saudi Arabia
| | | | - Monnera Saud Albagmi
- Department of Biochemistry, College of Science, King Saud University, Riyadh, 11451, Saudi Arabia
| | - Mohammed Arshad
- Dental Health Department, College of Applied Medical Sciences, King Saud University, Riyadh, Saudi Arabia
| | - Mohammad Tarique
- Department of Child Health, School of Medicine, University of Missouri, Columbia, MO 65201, USA
| | - Bilqees Bano
- Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, INDIA
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14
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Coccini T, Schicchi A, Locatelli CA, Caloni F, Negri S, Grignani E, De Simone U. Methylglyoxal-induced neurotoxic effects in primary neuronal-like cells transdifferentiated from human mesenchymal stem cells: Impact of low concentrations. J Appl Toxicol 2023; 43:1819-1839. [PMID: 37431083 DOI: 10.1002/jat.4515] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/16/2023] [Revised: 06/12/2023] [Accepted: 06/26/2023] [Indexed: 07/12/2023]
Abstract
In the last decades, advanced glycation end-products (AGEs) have aroused the interest of the scientific community due to the increasing evidence of their involvement in many pathophysiological processes including various neurological disorders and cognitive decline age related. Methylglyoxal (MG) is one of the reactive dicarbonyl precursors of AGEs, mainly generated as a by-product of glycolysis, whose accumulation induces neurotoxicity. In our study, MG cytotoxicity was evaluated employing a human stem cell-derived model, namely, neuron-like cells (hNLCs) transdifferentiated from mesenchymal stem/stromal cells, which served as a source of human based species-specific "healthy" cells. MG increased ROS production and induced the first characteristic apoptotic hallmarks already at low concentrations (≥10 μM), decreased the cell growth (≥5-10 μM) and viability (≥25 μM), altered Glo-1 and Glo-2 enzymes (≥25 μM), and markedly affected the neuronal markers MAP-2 and NSE causing their loss at low MG concentrations (≥10 μM). Morphological alterations started at 100 μM, followed by even more marked effects and cell death after few hours (5 h) from 200 μM MG addition. Substantially, most effects occurred as low as 10 μM, concentration much lower than that reported from previous observations using different in vitro cell-based models (e.g., human neuroblastoma cell lines, primary animal cells, and human iPSCs). Remarkably, this low effective concentration approaches the level range measured in biological samples of pathological subjects. The use of a suitable cellular model, that is, human primary neurons, can provide an additional valuable tool, mimicking better the physiological and biochemical properties of brain cells, in order to evaluate the mechanistic basis of molecular and cellular alterations in CNS.
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Affiliation(s)
- Teresa Coccini
- Laboratory of Clinical and Experimental Toxicology, and Pavia Poison Centre-National Toxicology Information Centre, Toxicology Unit, Istituti Clinici Scientifici Maugeri IRCCS, Pavia, Italy
| | - Azzurra Schicchi
- Laboratory of Clinical and Experimental Toxicology, and Pavia Poison Centre-National Toxicology Information Centre, Toxicology Unit, Istituti Clinici Scientifici Maugeri IRCCS, Pavia, Italy
| | - Carlo Alessandro Locatelli
- Laboratory of Clinical and Experimental Toxicology, and Pavia Poison Centre-National Toxicology Information Centre, Toxicology Unit, Istituti Clinici Scientifici Maugeri IRCCS, Pavia, Italy
| | - Francesca Caloni
- Dipartimento di Scienze e Politiche Ambientali (ESP), Università degli Studi di Milano, Milan, Italy
| | - Sara Negri
- Environmental Research Center, Istituti Clinici Scientifici Maugeri IRCCS, Pavia, Italy
| | - Elena Grignani
- Environmental Research Center, Istituti Clinici Scientifici Maugeri IRCCS, Pavia, Italy
| | - Uliana De Simone
- Laboratory of Clinical and Experimental Toxicology, and Pavia Poison Centre-National Toxicology Information Centre, Toxicology Unit, Istituti Clinici Scientifici Maugeri IRCCS, Pavia, Italy
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15
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Atawia RT, Batori R, Jordan CR, Kennard S, Antonova G, Bruder-Nascimento T, Mehta V, Saeed MI, Patel VS, Fukai T, Ushio-Fukai M, Huo Y, Fulton DJR, de Chantemèle EJB. Type 1 Diabetes Impairs Endothelium-Dependent Relaxation Via Increasing Endothelial Cell Glycolysis Through Advanced Glycation End Products, PFKFB3, and Nox1-Mediated Mechanisms. Hypertension 2023; 80:2059-2071. [PMID: 37729634 PMCID: PMC10514399 DOI: 10.1161/hypertensionaha.123.21341] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/16/2023] [Accepted: 08/02/2023] [Indexed: 09/22/2023]
Abstract
BACKGROUND Type 1 diabetes (T1D) is a major cause of endothelial dysfunction. Although cellular bioenergetics has been identified as a new regulator of vascular function, whether glycolysis, the primary bioenergetic pathway in endothelial cells (EC), regulates vascular tone and contributes to impaired endothelium-dependent relaxation (EDR) in T1D remains unknown. METHODS Experiments were conducted in Akita mice with intact or selective deficiency in EC PFKFB3 (6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3), the main regulator of glycolysis. Seahorse analyzer and myography were employed to measure glycolysis and mitochondrial respiration, and EDR, respectively, in aortic explants. EC PFKFB3 (Ad-PFKFB3) and glycolysis (Ad-GlycoHi) were increased in situ via adenoviral transduction. RESULTS T1D increased EC glycolysis and elevated EC expression of PFKFB3 and NADPH oxidase Nox1 (NADPH oxidase homolog 1). Functionally, pharmacological and genetic inhibition of PFKFB3 restored EDR in T1D, while in situ aorta EC transduction with Ad-PFKFB3 or Ad-GlycoHi reproduced the impaired EDR associated with T1D. Nox1 inhibition restored EDR in aortic rings from Akita mice, as well as in Ad-PFKFB3-transduced aorta EC and lactate-treated wild-type aortas. T1D increased the expression of the advanced glycation end product precursor methylglyoxal in the aortas. Exposure of the aortas to methylglyoxal impaired EDR, which was prevented by PFKFB3 inhibition. T1D and exposure to methylglyoxal increased EC expression of HIF1α (hypoxia-inducible factor 1α), whose inhibition blunted methylglyoxal-mediated EC PFKFB3 upregulation. CONCLUSIONS EC bioenergetics, namely glycolysis, is a new regulator of vasomotion and excess glycolysis, a novel mechanism of endothelial dysfunction in T1D. We introduce excess methylglyoxal, HIF1α, and PFKFB3 as major effectors in T1D-mediated increased EC glycolysis.
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Affiliation(s)
- Reem T. Atawia
- Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
- Department of Pharmacology and Toxicology, Faculty of Pharmacy, Ain Shams University, Abasia, Cairo, Egypt
| | - Robert Batori
- Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
| | - Coleton R. Jordan
- Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
| | - Simone Kennard
- Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
| | - Galina Antonova
- Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
| | | | - Vinay Mehta
- Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
| | - Muhammad I. Saeed
- Department of Surgery, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
| | - Vijay S Patel
- Department of Surgery, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
| | - Tohru Fukai
- Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
| | - Masuko Ushio-Fukai
- Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
| | - Yuqing Huo
- Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
| | - David JR Fulton
- Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, Georgia, USA
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16
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Baltazar P, de Melo Junior AF, Fonseca NM, Lança MB, Faria A, Sequeira CO, Teixeira-Santos L, Monteiro EC, Campos Pinheiro L, Calado J, Sousa C, Morello J, Pereira SA. Oxalate (dys)Metabolism: Person-to-Person Variability, Kidney and Cardiometabolic Toxicity. Genes (Basel) 2023; 14:1719. [PMID: 37761859 PMCID: PMC10530622 DOI: 10.3390/genes14091719] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/25/2023] [Revised: 08/11/2023] [Accepted: 08/16/2023] [Indexed: 09/29/2023] Open
Abstract
Oxalate is a metabolic end-product whose systemic concentrations are highly variable among individuals. Genetic (primary hyperoxaluria) and non-genetic (e.g., diet, microbiota, renal and metabolic disease) reasons underlie elevated plasma concentrations and tissue accumulation of oxalate, which is toxic to the body. A classic example is the triad of primary hyperoxaluria, nephrolithiasis, and kidney injury. Lessons learned from this example suggest further investigation of other putative factors associated with oxalate dysmetabolism, namely the identification of precursors (glyoxylate, aromatic amino acids, glyoxal and vitamin C), the regulation of the endogenous pathways that produce oxalate, or the microbiota's contribution to oxalate systemic availability. The association between secondary nephrolithiasis and cardiovascular and metabolic diseases (hypertension, type 2 diabetes, and obesity) inspired the authors to perform this comprehensive review about oxalate dysmetabolism and its relation to cardiometabolic toxicity. This perspective may offer something substantial that helps advance understanding of effective management and draws attention to the novel class of treatments available in clinical practice.
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Affiliation(s)
- Pedro Baltazar
- Centro Hospitalar Universitário de Lisboa Central, E.P.E, 1150-199 Lisboa, Portugal; (P.B.); (N.M.F.); (M.B.L.); (L.C.P.); (J.C.)
- iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal; (A.F.d.M.J.); (C.O.S.); (L.T.-S.); (E.C.M.); (C.S.); (J.M.)
- Centro Clínico Académico de Lisboa, 1159-056 Lisboa, Portugal
| | - Antonio Ferreira de Melo Junior
- iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal; (A.F.d.M.J.); (C.O.S.); (L.T.-S.); (E.C.M.); (C.S.); (J.M.)
- Centro Clínico Académico de Lisboa, 1159-056 Lisboa, Portugal
| | - Nuno Moreira Fonseca
- Centro Hospitalar Universitário de Lisboa Central, E.P.E, 1150-199 Lisboa, Portugal; (P.B.); (N.M.F.); (M.B.L.); (L.C.P.); (J.C.)
- Centro Clínico Académico de Lisboa, 1159-056 Lisboa, Portugal
| | - Miguel Brito Lança
- Centro Hospitalar Universitário de Lisboa Central, E.P.E, 1150-199 Lisboa, Portugal; (P.B.); (N.M.F.); (M.B.L.); (L.C.P.); (J.C.)
| | - Ana Faria
- CHRC, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal;
| | - Catarina O. Sequeira
- iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal; (A.F.d.M.J.); (C.O.S.); (L.T.-S.); (E.C.M.); (C.S.); (J.M.)
| | - Luísa Teixeira-Santos
- iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal; (A.F.d.M.J.); (C.O.S.); (L.T.-S.); (E.C.M.); (C.S.); (J.M.)
- Centro Clínico Académico de Lisboa, 1159-056 Lisboa, Portugal
| | - Emilia C. Monteiro
- iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal; (A.F.d.M.J.); (C.O.S.); (L.T.-S.); (E.C.M.); (C.S.); (J.M.)
- Centro Clínico Académico de Lisboa, 1159-056 Lisboa, Portugal
| | - Luís Campos Pinheiro
- Centro Hospitalar Universitário de Lisboa Central, E.P.E, 1150-199 Lisboa, Portugal; (P.B.); (N.M.F.); (M.B.L.); (L.C.P.); (J.C.)
- iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal; (A.F.d.M.J.); (C.O.S.); (L.T.-S.); (E.C.M.); (C.S.); (J.M.)
- Centro Clínico Académico de Lisboa, 1159-056 Lisboa, Portugal
| | - Joaquim Calado
- Centro Hospitalar Universitário de Lisboa Central, E.P.E, 1150-199 Lisboa, Portugal; (P.B.); (N.M.F.); (M.B.L.); (L.C.P.); (J.C.)
- Centro Clínico Académico de Lisboa, 1159-056 Lisboa, Portugal
- ToxOmics, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal
| | - Cátia Sousa
- iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal; (A.F.d.M.J.); (C.O.S.); (L.T.-S.); (E.C.M.); (C.S.); (J.M.)
- Centro Clínico Académico de Lisboa, 1159-056 Lisboa, Portugal
| | - Judit Morello
- iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal; (A.F.d.M.J.); (C.O.S.); (L.T.-S.); (E.C.M.); (C.S.); (J.M.)
| | - Sofia A. Pereira
- iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade NOVA de Lisboa, 1150-082 Lisboa, Portugal; (A.F.d.M.J.); (C.O.S.); (L.T.-S.); (E.C.M.); (C.S.); (J.M.)
- Centro Clínico Académico de Lisboa, 1159-056 Lisboa, Portugal
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Bednarska K, Fecka I, Scheijen JLJM, Ahles S, Vangrieken P, Schalkwijk CG. A Citrus and Pomegranate Complex Reduces Methylglyoxal in Healthy Elderly Subjects: Secondary Analysis of a Double-Blind Randomized Cross-Over Clinical Trial. Int J Mol Sci 2023; 24:13168. [PMID: 37685975 PMCID: PMC10488144 DOI: 10.3390/ijms241713168] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/29/2023] [Revised: 08/18/2023] [Accepted: 08/23/2023] [Indexed: 09/10/2023] Open
Abstract
Reactive α-dicarbonyls (α-DCs), such as methylglyoxal (MGO), glyoxal (GO), and 3-deoxyglucosone (3-DG), are potent precursors in the formation of advanced glycation end products (AGEs). In particular, MGO and MGO-derived AGEs are thought to be involved in the development of vascular complications in diabetes. Experimental studies showed that citrus and pomegranate polyphenols can scavenge α-DCs. Therefore, the aim of this study was to evaluate the effect of a citrus and pomegranate complex (CPC) on the α-DCs plasma levels in a double-blind, placebo-controlled cross-over trial, where thirty-six elderly subjects were enrolled. They received either 500 mg of Citrus sinensis peel extract and 200 mg of Punica granatum concentrate in CPC capsules or placebo capsules for 4 weeks, with a 4-week washout period in between. For the determination of α-DCs concentrations, liquid chromatography tandem mass spectrometry was used. Following four weeks of CPC supplementation, plasma levels of MGO decreased by 9.8% (-18.7 nmol/L; 95% CI: -36.7, -0.7 nmol/L; p = 0.042). Our findings suggest that CPC supplementation may represent a promising strategy for mitigating the conditions associated with MGO involvement. This study was registered on clinicaltrials.gov as NCT03781999.
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Affiliation(s)
- Katarzyna Bednarska
- Department of Pharmacognosy, Faculty of Pharmacy, Wroclaw Medical University, Borowska 211, 50-556 Wroclaw, Poland
| | - Izabela Fecka
- Department of Pharmacognosy, Faculty of Pharmacy, Wroclaw Medical University, Borowska 211, 50-556 Wroclaw, Poland
- The Committee on Therapeutics and Pharmaceutical Sciences, The Polish Academy of Sciences, Pl. Defilad 1, 00-901 Warsaw, Poland
| | - Jean L. J. M. Scheijen
- Department of Internal Medicine, Maastricht University Medical Center+, 6229 ER Maastricht, The Netherlands; (J.L.J.M.S.); (P.V.); (C.G.S.)
- CARIM School for Cardiovascular Diseases, Faculty of Health, Medicine and Life Sciences, Maastricht University, 6229 ER Maastricht, The Netherlands
| | - Sanne Ahles
- Department of Nutrition and Movement Sciences, School of Nutrition and Translational Research in Metabolism (NUTRIM), Maastricht University, 6229 ER Maastricht, The Netherlands;
- BioActor BV, 6229 GS Maastricht, The Netherlands
| | - Philippe Vangrieken
- Department of Internal Medicine, Maastricht University Medical Center+, 6229 ER Maastricht, The Netherlands; (J.L.J.M.S.); (P.V.); (C.G.S.)
- CARIM School for Cardiovascular Diseases, Faculty of Health, Medicine and Life Sciences, Maastricht University, 6229 ER Maastricht, The Netherlands
| | - Casper G. Schalkwijk
- Department of Internal Medicine, Maastricht University Medical Center+, 6229 ER Maastricht, The Netherlands; (J.L.J.M.S.); (P.V.); (C.G.S.)
- CARIM School for Cardiovascular Diseases, Faculty of Health, Medicine and Life Sciences, Maastricht University, 6229 ER Maastricht, The Netherlands
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Remund B, Yilmaz B, Sokollik C. D-Lactate: Implications for Gastrointestinal Diseases. CHILDREN (BASEL, SWITZERLAND) 2023; 10:945. [PMID: 37371177 DOI: 10.3390/children10060945] [Citation(s) in RCA: 9] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/30/2023] [Revised: 05/19/2023] [Accepted: 05/24/2023] [Indexed: 06/29/2023]
Abstract
D-lactate is produced in very low amounts in human tissues. However, certain bacteria in the human intestine produce D-lactate. In some gastrointestinal diseases, increased bacterial D-lactate production and uptake from the gut into the bloodstream take place. In its extreme, excessive accumulation of D-lactate in humans can lead to potentially life-threatening D-lactic acidosis. This metabolic phenomenon is well described in pediatric patients with short bowel syndrome. Less is known about a subclinical rise in D-lactate. We discuss in this review the pathophysiology of D-lactate in the human body. We cover D-lactic acidosis in patients with short bowel syndrome as well as subclinical elevations of D-lactate in other diseases affecting the gastrointestinal tract. Furthermore, we argue for the potential of D-lactate as a marker of intestinal barrier integrity in the context of dysbiosis. Subsequently, we conclude that there is a research need to establish D-lactate as a minimally invasive biomarker in gastrointestinal diseases.
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Affiliation(s)
- Barblin Remund
- Division of Paediatric Gastroenterology, Hepatology and Nutrition, Department of Paediatrics, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, Switzerland
| | - Bahtiyar Yilmaz
- Department of Visceral Surgery and Medicine, Bern University Hospital, University of Bern, 3010 Bern, Switzerland
- Maurice Müller Laboratories, Department for Biomedical Research, University of Bern, 3008 Bern, Switzerland
| | - Christiane Sokollik
- Division of Paediatric Gastroenterology, Hepatology and Nutrition, Department of Paediatrics, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, Switzerland
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19
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Chegão A, Vicente Miranda H. Unveiling new secrets in Parkinson's disease: The glycatome. Behav Brain Res 2023; 442:114309. [PMID: 36706808 DOI: 10.1016/j.bbr.2023.114309] [Citation(s) in RCA: 6] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/15/2022] [Revised: 01/04/2023] [Accepted: 01/19/2023] [Indexed: 01/25/2023]
Abstract
We are witnessing a considerable increase in the incidence of Parkinson's disease (PD), which may be due to the general ageing of the population. While there is a plethora of therapeutic strategies for this disease, they still fail to arrest disease progression as they do not target and prevent the neurodegenerative process. The identification of disease-causing mutations allowed researchers to better dissect the underlying causes of this disease, highlighting, for example, the pathogenic role of alpha-synuclein. However, most PD cases are sporadic, which is making it hard to unveil the major causative mechanisms of this disease. In the recent years, epidemiological evidence suggest that type-2 diabetes mellitus (T2DM) individuals have higher risk and worst outcomes of PD, allowing to raise the hypothesis that some dysregulated processes in T2DM may contribute or even trigger the neurodegenerative process in PD. One major consequence of T2DM is the unprogrammed reaction between sugars, increased in T2DM, and proteins, a reaction named glycation. Pre-clinical reports show that alpha-synuclein is a target of glycation, and glycation potentiates its pathogenicity which contributes for the neurodegenerative process. Moreover, it triggers, anticipates, or aggravates several PD-like motor and non-motor complications. A given profile of proteins are differently glycated in diseased conditions, altering the brain proteome and leading to brain dysfunction and neurodegeneration. Herein we coin the term Glycatome as the profile of glycated proteins. In this review we report on the mechanisms underlying the association between T2DM and PD, with particular focus on the impact of protein glycation.
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Affiliation(s)
- Ana Chegão
- iNOVA4Health, NOVA Medical School, NMS, Universidade NOVA de Lisboa, Lisboa, Portugal
| | - Hugo Vicente Miranda
- iNOVA4Health, NOVA Medical School, NMS, Universidade NOVA de Lisboa, Lisboa, Portugal.
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20
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Hagenhaus V, Gorenflos López JL, Rosenstengel R, Neu C, Hackenberger CPR, Celik A, Weinert K, Nguyen MB, Bork K, Horstkorte R, Gesper A. Glycation Interferes with the Activity of the Bi-Functional UDP- N-Acetylglucosamine 2-Epimerase/ N-Acetyl-mannosamine Kinase (GNE). Biomolecules 2023; 13:biom13030422. [PMID: 36979358 PMCID: PMC10046061 DOI: 10.3390/biom13030422] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/23/2022] [Revised: 02/17/2023] [Accepted: 02/20/2023] [Indexed: 03/30/2023] Open
Abstract
Mutations in the gene coding for the bi-functional UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE), the key enzyme of the sialic acid biosynthesis, are responsible for autosomal-recessive GNE myopathy (GNEM). GNEM is an adult-onset disease with a yet unknown exact pathophysiology. Since the protein appears to work adequately for a certain period of time even though the mutation is already present, other effects appear to influence the onset and progression of the disease. In this study, we want to investigate whether the late onset of GNEM is based on an age-related effect, e.g., the accumulation of post-translational modifications (PTMs). Furthermore, we also want to investigate what effect on the enzyme activity such an accumulation would have. We will particularly focus on glycation, which is a PTM through non-enzymatic reactions between the carbonyl groups (e.g., of methylglyoxal (MGO) or glyoxal (GO)) with amino groups of proteins or other biomolecules. It is already known that the levels of both MGO and GO increase with age. For our investigations, we express each domain of the GNE separately, treat them with one of the glycation agents, and determine their activity. We demonstrate that the enzymatic activity of the N-acetylmannosamine kinase (GNE-kinase domain) decreases dramatically after glycation with MGO or GO-with a remaining activity of 13% ± 5% (5 mM MGO) and 22% ± 4% (5 mM GO). Whereas the activity of the UDP-N-acetylglucosamine 2-epimerase (GNE-epimerase domain) is only slightly reduced after glycation-with a remaining activity of 60% ± 8% (5 mM MGO) and 63% ± 5% (5 mM GO).
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Affiliation(s)
- Vanessa Hagenhaus
- Institute for Physiological Chemistry, Medical Faculty, Martin-Luther-University Halle-Wittenberg, 06114 Halle, Germany
| | - Jacob L Gorenflos López
- Leibniz-Forschungsinstitut für Molekulare Pharmakologie im Forschungsverbund Berlin e.V. (FMP), Campus Berlin-Buch, Robert-Roessle-Str. 10, 13125 Berlin, Germany
- Institut für Chemie, Humboldt Universität zu Berlin, Brook-Taylor-Str. 2, 12489 Berlin, Germany
| | - Rebecca Rosenstengel
- Institute for Physiological Chemistry, Medical Faculty, Martin-Luther-University Halle-Wittenberg, 06114 Halle, Germany
| | - Carolin Neu
- Institute for Physiological Chemistry, Medical Faculty, Martin-Luther-University Halle-Wittenberg, 06114 Halle, Germany
| | - Christian P R Hackenberger
- Leibniz-Forschungsinstitut für Molekulare Pharmakologie im Forschungsverbund Berlin e.V. (FMP), Campus Berlin-Buch, Robert-Roessle-Str. 10, 13125 Berlin, Germany
- Institut für Chemie, Humboldt Universität zu Berlin, Brook-Taylor-Str. 2, 12489 Berlin, Germany
| | - Arif Celik
- Leibniz-Forschungsinstitut für Molekulare Pharmakologie im Forschungsverbund Berlin e.V. (FMP), Campus Berlin-Buch, Robert-Roessle-Str. 10, 13125 Berlin, Germany
- Institut für Chemie, Humboldt Universität zu Berlin, Brook-Taylor-Str. 2, 12489 Berlin, Germany
| | - Klara Weinert
- Institute for Physiological Chemistry, Medical Faculty, Martin-Luther-University Halle-Wittenberg, 06114 Halle, Germany
| | - Mai-Binh Nguyen
- Institute for Physiological Chemistry, Medical Faculty, Martin-Luther-University Halle-Wittenberg, 06114 Halle, Germany
| | - Kaya Bork
- Institute for Physiological Chemistry, Medical Faculty, Martin-Luther-University Halle-Wittenberg, 06114 Halle, Germany
| | - Rüdiger Horstkorte
- Institute for Physiological Chemistry, Medical Faculty, Martin-Luther-University Halle-Wittenberg, 06114 Halle, Germany
| | - Astrid Gesper
- Institute for Physiological Chemistry, Medical Faculty, Martin-Luther-University Halle-Wittenberg, 06114 Halle, Germany
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21
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Lai SWT, Lopez Gonzalez EDJ, Zoukari T, Ki P, Shuck SC. Methylglyoxal and Its Adducts: Induction, Repair, and Association with Disease. Chem Res Toxicol 2022; 35:1720-1746. [PMID: 36197742 PMCID: PMC9580021 DOI: 10.1021/acs.chemrestox.2c00160] [Citation(s) in RCA: 68] [Impact Index Per Article: 22.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/17/2023]
Abstract
Metabolism is an essential part of life that provides energy for cell growth. During metabolic flux, reactive electrophiles are produced that covalently modify macromolecules, leading to detrimental cellular effects. Methylglyoxal (MG) is an abundant electrophile formed from lipid, protein, and glucose metabolism at intracellular levels of 1-4 μM. MG covalently modifies DNA, RNA, and protein, forming advanced glycation end products (MG-AGEs). MG and MG-AGEs are associated with the onset and progression of many pathologies including diabetes, cancer, and liver and kidney disease. Regulating MG and MG-AGEs is a potential strategy to prevent disease, and they may also have utility as biomarkers to predict disease risk, onset, and progression. Here, we review recent advances and knowledge surrounding MG, including its production and elimination, mechanisms of MG-AGEs formation, the physiological impact of MG and MG-AGEs in disease onset and progression, and the latter in the context of its receptor RAGE. We also discuss methods for measuring MG and MG-AGEs and their clinical application as prognostic biomarkers to allow for early detection and intervention prior to disease onset. Finally, we consider relevant clinical applications and current therapeutic strategies aimed at targeting MG, MG-AGEs, and RAGE to ultimately improve patient outcomes.
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Affiliation(s)
- Seigmund Wai Tsuen Lai
- Department of Diabetes and Cancer Metabolism, Arthur Riggs Diabetes and Metabolism Research Institute, City of Hope Comprehensive Cancer Center, Duarte, California 91010, United States
| | - Edwin De Jesus Lopez Gonzalez
- Department of Diabetes and Cancer Metabolism, Arthur Riggs Diabetes and Metabolism Research Institute, City of Hope Comprehensive Cancer Center, Duarte, California 91010, United States
| | - Tala Zoukari
- Department of Diabetes and Cancer Metabolism, Arthur Riggs Diabetes and Metabolism Research Institute, City of Hope Comprehensive Cancer Center, Duarte, California 91010, United States
| | - Priscilla Ki
- Department of Diabetes and Cancer Metabolism, Arthur Riggs Diabetes and Metabolism Research Institute, City of Hope Comprehensive Cancer Center, Duarte, California 91010, United States
| | - Sarah C Shuck
- Department of Diabetes and Cancer Metabolism, Arthur Riggs Diabetes and Metabolism Research Institute, City of Hope Comprehensive Cancer Center, Duarte, California 91010, United States
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22
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Martin-Morales A, Arakawa T, Sato M, Matsumura Y, Mano-Usui F, Ikeda K, Inagaki N, Sato K. Development of a Method for Quantitation of Glyceraldehyde in Various Body Compartments of Rodents and Humans. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 2021; 69:13246-13254. [PMID: 34702032 DOI: 10.1021/acs.jafc.1c03177] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/13/2023]
Abstract
There is limited information available about the physiological content of glyceraldehyde, a precursor of toxic advanced glycation end products. The conventional derivatization method for aldoses using 1-phenyl-3-methyl-5-pyrazolone did not allow reproducible quantification of glyceraldehyde due to the instability of glyceraldehyde compared to other aldoses. We optimized the derivatization condition to achieve high and reproducible recovery of derivatives for liquid chromatography tandem mass spectrometry quantification. Based on the stability of glyceraldehyde during sample preparation and high recovery of spiked standard, the present method provides reproducible quantification of glyceraldehyde in the body. The glyceraldehyde contents in fasting conditions in the rodent liver (mice: 50.0 ± 3.9 nmol/g; rats: 35.5 ± 4.9 nmol/g) were higher than those in plasma (9.4 ± 1.7 and 7.2 ± 1.2 nmol/mL). The liver glyceraldehyde levels significantly increased after food consumption (p < 0.05) but remained constant in the plasma. High fat diet feeding significantly increased plasma glyceraldehyde levels in mice (p < 0.005). In healthy human volunteers, the plasma glyceraldehyde levels remained unchanged after the consumption of steamed rice. In patients with type 2 diabetes, the plasma glyceraldehyde level was positively correlated with the plasma glucose level (r = 0.84; p < 0.0001).
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Affiliation(s)
- Agustin Martin-Morales
- Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Kyoto 606 8054, Japan
| | - Takanori Arakawa
- Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Kyoto 606 8054, Japan
| | - Mona Sato
- Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Kyoto 606 8054, Japan
| | - Yasuki Matsumura
- Division of Agronomy and Horticultural Science, Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611 0011, Japan
| | - Fumika Mano-Usui
- Department of Diabetes, Endocrinology and Nutrition, Graduate School of Medicine, Kyoto University, 54 Shogoin Kawahara-cho, Kyoto 606 8507, Japan
| | - Kaori Ikeda
- Department of Diabetes, Endocrinology and Nutrition, Graduate School of Medicine, Kyoto University, 54 Shogoin Kawahara-cho, Kyoto 606 8507, Japan
| | - Nobuya Inagaki
- Department of Diabetes, Endocrinology and Nutrition, Graduate School of Medicine, Kyoto University, 54 Shogoin Kawahara-cho, Kyoto 606 8507, Japan
| | - Kenji Sato
- Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Kyoto 606 8054, Japan
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23
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Zhao Y, Tang Y, Sang S. Dietary Quercetin Reduces Plasma and Tissue Methylglyoxal and Advanced Glycation End Products in Healthy Mice Treated with Methylglyoxal. J Nutr 2021; 151:2601-2609. [PMID: 34091674 DOI: 10.1093/jn/nxab176] [Citation(s) in RCA: 12] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/22/2021] [Revised: 03/15/2021] [Accepted: 05/11/2021] [Indexed: 01/17/2023] Open
Abstract
BACKGROUND Methylglyoxal (MGO), a precursor of advanced glycation end products (AGEs), has been linked to AGEs-associated diseases. OBJECTIVES This study investigated the efficacy and mechanisms of dietary quercetin in decreasing plasma and tissue concentrations of MGO and AGEs in MGO-administered mice. METHODS Male, 6-wk-old CD-1 mice were administered AIN-93G diet and water (Con) or 0.12% MGO in water (MGO) or MGO plus 0.2% (0.2Q) dietary quercetin for 1 wk (n = 5) (experiment 1), and water (Con), 0.12% MGO (MGO), or MGO plus 0.1% (0.1Q), 0.2% (0.2Q), or 0.4% (0.4Q) dietary quercetin for 6 wk (n = 10) (experiment 2). The plasma, kidney, and liver concentrations of MGO, quercetin, and isorhamnetin and their trapping adducts with MGO were determined by LC-MS, and AGE concentrations were measured by the fluorescent method. Furthermore, the expressions of glyoxalase I/II (GLO I/II) and aldose reductase (AR), MGO detoxification enzymes, were determined by Western blot. One-factor ANOVA and post hoc Dunnett's or Tukey's test were used to analyze the data. RESULTS After 1 wk of treatment, the MGO concentrations in plasma (20.2%) and kidney (29.9%) in 0.2Q mice were significantly lower than those in MGO mice. After 6 wk of treatment, the concentrations of MGO in the plasma (14.7-18.6%), kidney (20-20.8%), liver (15.4-18.6%), and tissue AGEs (28-36.8%) in 0.1Q, 0.2Q, and 0.4Q mice were significantly lower than those in MGO mice. The plasma concentrations of quercetin, isorhamnetin, and their MGO adducts were dose-dependently increased after quercetin administration. In addition, after 6 wk of quercetin administration, the expressions of GLO I/II and AR in the liver and kidney were significantly upregulated to promote MGO detoxification compared with MGO-treated mice. CONCLUSIONS Quercetin reduced plasma and tissue MGO concentrations and inhibited AGE formation by trapping MGO and regulating the MGO detoxification systems in MGO-administered healthy mice.
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Affiliation(s)
- Yantao Zhao
- Laboratory for Functional Foods and Human Health, Center for Excellence in Post-Harvest Technologies, North Carolina Agricultural and Technical State University, North Carolina Research Campus, Kannapolis, NC, USA
| | - Yao Tang
- Laboratory for Functional Foods and Human Health, Center for Excellence in Post-Harvest Technologies, North Carolina Agricultural and Technical State University, North Carolina Research Campus, Kannapolis, NC, USA
| | - Shengmin Sang
- Laboratory for Functional Foods and Human Health, Center for Excellence in Post-Harvest Technologies, North Carolina Agricultural and Technical State University, North Carolina Research Campus, Kannapolis, NC, USA
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24
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Phenethylamine in chlorella alleviates high-fat diet-induced mouse liver damage by regulating generation of methylglyoxal. NPJ Sci Food 2021; 5:22. [PMID: 34301957 PMCID: PMC8302609 DOI: 10.1038/s41538-021-00105-3] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/04/2021] [Accepted: 07/06/2021] [Indexed: 01/27/2023] Open
Abstract
This study examined the effects of oral administration of water extract of chlorella (WEC) (100 mg/kg bodyweight) and phenethylamine (10 μg/kg bodyweight) on high-fat diet (HFD)-induced liver damage in mice. Phenethylamine significantly mitigated HFD-induced lipid oxidation (generation of malondialdehyde) and liver damage without markedly decreasing hepatic lipid accumulation. WEC exerted similar effects although with decreased efficacy. In addition, WEC and phenethylamine decreased the methylglyoxal levels and increased the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein levels in the liver. Methylglyoxal is generated from substrates of GAPDH, dihydroxyacetone phosphate and glyceraldehyde 3-phosphate. These facts indicate that methylglyoxal triggers oxidation of accumulated lipid, which generates malondialdehyde and consequently induces liver damage. Suppression of generation of toxic aldehydes by WEC and phenethylamine was also confirmed by maintaining hepatic cysteine, highly reactive to aldehydes. Thus, trace amounts of phenethylamine alleviate HFD-induced liver damage by regulating methylglyoxal via increase of GAPDH.
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25
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Briens JM, Subramaniam M, Kilgour A, Loewen ME, Desai KM, Adolphe JL, Zatti KM, Drew MD, Weber LP. Glycemic, insulinemic and methylglyoxal postprandial responses to starches alone or in whole diets in dogs versus cats: Relating the concept of glycemic index to metabolic responses and gene expression. Comp Biochem Physiol A Mol Integr Physiol 2021; 257:110973. [PMID: 33933629 DOI: 10.1016/j.cbpa.2021.110973] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/12/2020] [Revised: 04/26/2021] [Accepted: 04/27/2021] [Indexed: 10/21/2022]
Abstract
Species differences between domestic cats (Felis catus) and dogs (Canis familiaris) has led to differences in their ability to digest, absorb and metabolize carbohydrates through poorly characterized mechanisms. The current study aimed to first examine biopsied small intestine, pancreas, liver and skeletal muscle from laboratory beagles and domestic cats for mRNA expression of key enzymes involved in starch digestion (amylase), glucose transport (sodium-dependent SGLTs and -independent glucose transporters, GLUT) and glucose metabolism (hexokinase and glucokinase). Cats had lower mRNA expression of most genes examined in almost all tissues compared to dogs (p < 0.05). Next, postprandial glucose, insulin, methylglyoxal (a toxic glucose metabolite) and d-lactate (metabolite of methylglyoxal) after single feedings of different starch sources were tested in fasted dogs and cats. After feeding pure glucose, peak postprandial blood glucose and methylglyoxal were surprisingly similar between dogs and cats, except cats had a longer time to peak and a greater area under the curve consistent with lower glycolytic enzyme expression. After feeding starches or whole diets to dogs, postprandial glycemic response, glycemic index, insulin, methylglyoxal and d-lactate followed reported glycemic index trends in humans. In contrast, cats showed very low to negligible postprandial glycemic responses and low insulin after feeding different starch sources, but not whole diets, with no relationship to methylglyoxal or d-lactate. Thus, the concept of glycemic index appears valid in dogs, but not cats. Differences in amylase, glucose transporters, and glycolytic enzymes are consistent with species differences in starch and glucose handling between cats and dogs.
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Affiliation(s)
- Jennifer M Briens
- Toxicology Graduate Program, University of Saskatchewan, Saskatoon, SK S7N 5B3, Canada
| | - Marina Subramaniam
- Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, SK S7N 5B4, Canada
| | - Alyssa Kilgour
- Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, SK S7N 5B4, Canada
| | - Matthew E Loewen
- Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, SK S7N 5B4, Canada
| | - Kaushik M Desai
- Pharmacology & Physiology, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada
| | - Jennifer L Adolphe
- Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, SK S7N 5B4, Canada
| | - Kyla M Zatti
- Animal & Poultry Science, University of Saskatchewan, Saskatoon, SK S7N 5A8, Canada
| | - Murray D Drew
- Animal & Poultry Science, University of Saskatchewan, Saskatoon, SK S7N 5A8, Canada
| | - Lynn P Weber
- Toxicology Graduate Program, University of Saskatchewan, Saskatoon, SK S7N 5B3, Canada; Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, SK S7N 5B4, Canada.
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26
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Ovbude ST, Tao P, Li Z, Hage DS. High-Performance affinity chromatographic studies of repaglinide and nateglinide interactions with normal and glyoxal- or methylglyoxal-modified human albumin serum. J Pharm Biomed Anal 2021; 201:114097. [PMID: 33933705 DOI: 10.1016/j.jpba.2021.114097] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/18/2021] [Revised: 04/17/2021] [Accepted: 04/19/2021] [Indexed: 11/16/2022]
Abstract
During diabetes human serum albumin (HSA), an important drug transport protein, can be modified by agents such as glyoxal (Go) and methylglyoxal (MGo) to form advanced glycation end-products. High-performance affinity microcolumns and zonal elution competition studies were used to compare interactions by the anti-diabetic drugs repaglinide and nateglinide with normal and Go- or MGo-modified HSA at Sudlow sites I and II of this protein. Both drugs had their strongest binding at Sudlow site II for the normal and modified forms of HSA. The association equilibrium constants at this site for repaglinide and nateglinide with normal HSA were 6.1 (± 0.2) × 104 M-1 and 7.1 (± 0.8) × 105 M-1, respectively, at pH 7.4 and 37⁰C; these values increased by up to 3.6-fold for repaglinide and decreased by up to 45-55 % for nateglinide when HSA was modified by Go or MGo at levels seen in prediabetes or diabetes. Both drugs were also found to bind at Sudlow site I, with association equilibrium constants at this site on normal HSA of 4.2 (± 0.3) × 104 M-1 for repaglinide and 5.0 (± 0.1) × 104 M-1 for nateglinide. The binding strength for repaglinide at Sudlow site I increased by 1.3- to 1.7-fold with the Go-modified HSA and decreased slightly (i.e., up to 19 %) for the MGo-modified HSA, while nateglinide showed only a small or insignificant change in binding with the same modified HSA samples. These results indicated that binding by repaglinide and nateglinide with HSA can be altered significantly by modification of this protein with Go or MGo, making these modifications of potential interest in the treatment of patients with these drugs during diabetes.
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Affiliation(s)
- Susan T Ovbude
- Chemistry Department, University of Nebraska, Lincoln, NE, 68588, USA
| | - Pingyang Tao
- Chemistry Department, University of Nebraska, Lincoln, NE, 68588, USA
| | - Zhao Li
- Chemistry Department, University of Nebraska, Lincoln, NE, 68588, USA
| | - David S Hage
- Chemistry Department, University of Nebraska, Lincoln, NE, 68588, USA.
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Guillon C, Ferraro S, Clément S, Bouschbacher M, Sigaudo-Roussel D, Bonod C. Glycation by glyoxal leads to profound changes in the behavior of dermal fibroblasts. BMJ Open Diabetes Res Care 2021; 9:9/1/e002091. [PMID: 33903117 PMCID: PMC8076933 DOI: 10.1136/bmjdrc-2020-002091] [Citation(s) in RCA: 14] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/22/2020] [Revised: 03/17/2021] [Accepted: 03/21/2021] [Indexed: 12/23/2022] Open
Abstract
INTRODUCTION Diabetes is a worldwide health problem that is associated with severe complications. Advanced Glycation End products (AGEs) such as Nε-(carboxymethyl)lysine, which result from chronic hyperglycemia, accumulate in the skin of patients with diabetes. The effect of AGEs on fibroblast functionality and their impact on wound healing are still poorly understood. RESEARCH DESIGN AND METHODS To investigate this, we treated cultured human fibroblasts with 0.6 mM glyoxal to induce acute glycation. The behavior of fibroblasts was analyzed by time-lapse monolayer wounding healing assay, seahorse technology and atomic force microscopy. Production of extracellular matrix was studied by transmission electronic microscopy and western blot. Lipid metabolism was investigated by staining of lipid droplets (LDs) with BODIPY 493/503. RESULTS We found that the proliferative and migratory capacities of the cells were greatly reduced by glycation, which could be explained by an increase in fibroblast tensile strength. Measurement of the cellular energy balance did not indicate that there was a change in the rate of oxygen consumption of the fibroblasts. Assessment of collagen I revealed that glyoxal did not influence type I collagen secretion although it did disrupt collagen I maturation and it prevented its deposition in the extracellular matrix. We noted a pronounced increase in the number of LDs after glyoxal treatment. AMPK phosphorylation was reduced by glyoxal treatment but it was not responsible for the accumulation of LDs. CONCLUSION Glyoxal promotes a change in fibroblast behavior in favor of lipogenic activity that could be involved in delaying wound healing.
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Affiliation(s)
- Cécile Guillon
- Urgo Research Innovation and Development, Chenôve, France
- Université de Lyon, Université Claude Bernard Lyon 1, CNRS, LBTI UMR 5305, Lyon, France
| | - Sandra Ferraro
- Université de Lyon, Université Claude Bernard Lyon 1, CNRS, LBTI UMR 5305, Lyon, France
| | - Sophie Clément
- Urgo Research Innovation and Development, Chenôve, France
| | | | | | - Christelle Bonod
- Université de Lyon, Université Claude Bernard Lyon 1, CNRS, LBTI UMR 5305, Lyon, France
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Zhao Q, Li H, Chen H, Wu C, Ei-Seedi H, Xu X, Du M. High throughput analysis and quantitation of α-dicarbonyls in biofluid by plasmonic nanoshells enhanced laser desorption/ionization mass spectrometry. JOURNAL OF HAZARDOUS MATERIALS 2021; 403:123580. [PMID: 33264850 DOI: 10.1016/j.jhazmat.2020.123580] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/29/2020] [Revised: 07/22/2020] [Accepted: 07/23/2020] [Indexed: 06/12/2023]
Abstract
Advanced analytical platforms are required for accurate detection and quantification of small molecular substances exhibiting certain toxicity. Small molecules detection in complex biological fluids are challenged by the complexity of the samples and the low throughput of the existing methods. In the present study, to detect a batch of samples (50) in 1 h, the plasmonic nanoshell enhanced matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) was tested. The limit of quantification (LOQ) was determined as 0.01 μg/mL (for α-dicarbonyl compounds) by vortex-assisted liquid-liquid microextraction (VALLME). The developed method can be adopted to study the high-throughput metabolomics and employed for clinical precision diagnosis with MALDI-TOF MS.
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Affiliation(s)
- Qiyue Zhao
- National Engineering Research Center of Seafood, Collaborative Innovation Center of Provincial and Ministerial Co-Construction for Seafood Deep Processing, School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, People's Republic of China
| | - Han Li
- National Engineering Research Center of Seafood, Collaborative Innovation Center of Provincial and Ministerial Co-Construction for Seafood Deep Processing, School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, People's Republic of China
| | - Hui Chen
- National Engineering Research Center of Seafood, Collaborative Innovation Center of Provincial and Ministerial Co-Construction for Seafood Deep Processing, School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, People's Republic of China
| | - Chao Wu
- National Engineering Research Center of Seafood, Collaborative Innovation Center of Provincial and Ministerial Co-Construction for Seafood Deep Processing, School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, People's Republic of China
| | - Hesham Ei-Seedi
- Department of Medicinal Chemistry, Uppsala University, Biomedical Centre, Uppsala, Sweden
| | - Xianbing Xu
- National Engineering Research Center of Seafood, Collaborative Innovation Center of Provincial and Ministerial Co-Construction for Seafood Deep Processing, School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, People's Republic of China.
| | - Ming Du
- National Engineering Research Center of Seafood, Collaborative Innovation Center of Provincial and Ministerial Co-Construction for Seafood Deep Processing, School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, People's Republic of China.
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Rezaei M, Kalantari H, Mehrzadi S, Goudarzi M. Synergy Effects of Metformin and Berberine on Glyoxal-induced Carbonyl Stress in Isolated Rat Liver Mitochondria. CURRENT DRUG THERAPY 2021. [DOI: 10.2174/1574885515666200214122055] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
Objective:
Carbonyl stress, resulting from toxic effects of alpha-dicarbonyls such as
glyoxal (GO), plays an important role in mitochondrial dysfunction and subsequent development of
diabetic complications. This study evaluated the ability of metformin (MET), berberine (BBR), and
their combination to prevent GO-induced carbonyl stress in isolated rat liver mitochondria.
Methods:
Mitochondria (0.5 mg protein/mL) were isolated from the Wistar rat liver and incubated
with various concentrations of GO (1, 2.5, 5, 7.5, and 10 mM) for 30 minutes and IC50 for GO was
calculated. The suspensions of mitochondria were incubated with various concentrations of MET
(2.5, 5, 10, and 20 mM) or BBR (2.5, 5, 10, and 20 μM) for 30 min and then GO in a dose of IC50
at 37 ºC for 30 min. Mitochondrial complex II activity, mitochondrial membrane potential (MMP),
MDA level, reactive oxygen species (ROS) formation, reduced glutathione (GSH) content, and
protein carbonylation were assessed. The combination index and isobologram of MET and BBR on
GO toxicity were calculated.
Results:
IC50 of GO was assigned approximately 3 mM. GO disrupted the electron transfer chain
and significantly increased mitochondrial ROS formation, protein carbonylation, and MDA level.
GO decreased mitochondrial viability, MMP, and GSH content. Pre-treatment with MET and BBR
could potentially reverse GO-induced deleterious effects in a concentration-dependent manner.
Results of the drug combination indicated that CI for Fa 0.5 (Effect 50 %) was 0.83.
Conclusion:
These results suggest that BBR in combination with MET has a moderate synergistic
effect on GO-induced carbonyl stress in isolated rat liver mitochondria.
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Affiliation(s)
- Mohsen Rezaei
- Medicinal Plant Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Heibatullah Kalantari
- Medicinal Plant Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Saeed Mehrzadi
- Razi Drug Research Center, Iran University of Medical Sciences, Tehran, Iran
| | - Mehdi Goudarzi
- Medicinal Plant Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
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30
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Bednarska K, Kuś P, Fecka I. Investigation of the Phytochemical Composition, Antioxidant Activity, and Methylglyoxal Trapping Effect of Galega officinalis L. Herb In Vitro. Molecules 2020; 25:E5810. [PMID: 33317096 PMCID: PMC7764533 DOI: 10.3390/molecules25245810] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/17/2020] [Revised: 12/04/2020] [Accepted: 12/07/2020] [Indexed: 12/28/2022] Open
Abstract
Galega officinalis L. has been known for centuries as an herbal medicine used to alleviate the symptoms of diabetes, but its comprehensive chemical composition and pharmacological activity are still insufficiently known. The current study involved the qualitative and quantitative phytochemical analysis and in vitro evaluation of the antioxidative and methylglyoxal (MGO) trapping properties of galega herb. Ultra high-performance liquid chromatography coupled with both the electrospray ionization mass spectrometer and diode-array detector (UHPLC-ESI-MS and UHPLC-DAD) were used to investigate the composition and evaluate the anti-MGO capability of extracts and their components. Hot water and aqueous methanol extracts, as well as individual compounds representing phytochemical groups, were also assessed for antioxidant activity using DPPH (2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) assays. Quercetin and metformin were used as a positive control. We confirmed the presence of tricyclic quinazoline alkaloids, guanidines, flavonoids, and hydroxycinnamic acids (HCAs) in galega extracts. The polyphenolic fraction was dominated by mono-, di-, and triglycosylated flavonols, as well as monocaffeoylhexaric acids. The in vitro tests indicated which G. officinalis components exhibit beneficial antioxidative and MGO trapping effects. For galega extracts, flavonols, and HCAs, a potent antiradical activity was observed. The ability to trap MGO was noted for guanidines and flavonoids, whereas HCA esters and quinazoline alkaloids were ineffective. The formation of mono-MGO adducts of galegine, hydroxygalegine, and rutin in the examined water infusion was observed.
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Affiliation(s)
- Katarzyna Bednarska
- Department of Pharmacognosy and Herbal Medicines, Faculty of Pharmacy, Wroclaw Medical University, ul. Borowska 211, 50-556 Wroclaw, Poland; (P.K.); (I.F.)
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31
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Dicarbonyl derived post-translational modifications: chemistry bridging biology and aging-related disease. Essays Biochem 2020; 64:97-110. [PMID: 31939602 DOI: 10.1042/ebc20190057] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2019] [Revised: 12/17/2019] [Accepted: 12/19/2019] [Indexed: 01/17/2023]
Abstract
In living systems, nucleophilic amino acid residues are prone to non-enzymatic post-translational modification by electrophiles. α-Dicarbonyl compounds are a special type of electrophiles that can react irreversibly with lysine, arginine, and cysteine residues via complex mechanisms to form post-translational modifications known as advanced glycation end-products (AGEs). Glyoxal, methylglyoxal, and 3-deoxyglucosone are the major endogenous dicarbonyls, with methylglyoxal being the most well-studied. There are several routes that lead to the formation of dicarbonyl compounds, most originating from glucose and glucose metabolism, such as the non-enzymatic decomposition of glycolytic intermediates and fructosyl amines. Although dicarbonyls are removed continuously mainly via the glyoxalase system, several conditions lead to an increase in dicarbonyl concentration and thereby AGE formation. AGEs have been implicated in diabetes and aging-related diseases, and for this reason the elucidation of their structure as well as protein targets is of great interest. Though the dicarbonyls and reactive protein side chains are of relatively simple nature, the structures of the adducts as well as their mechanism of formation are not that trivial. Furthermore, detection of sites of modification can be demanding and current best practices rely on either direct mass spectrometry or various methods of enrichment based on antibodies or click chemistry followed by mass spectrometry. Future research into the structure of these adducts and protein targets of dicarbonyl compounds may improve the understanding of how the mechanisms of diabetes and aging-related physiological damage occur.
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Kim D, Kim KA, Kim JH, Kim EH, Bae ON. Methylglyoxal-Induced Dysfunction in Brain Endothelial Cells via the Suppression of Akt/HIF-1α Pathway and Activation of Mitophagy Associated with Increased Reactive Oxygen Species. Antioxidants (Basel) 2020; 9:antiox9090820. [PMID: 32899154 PMCID: PMC7554889 DOI: 10.3390/antiox9090820] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/09/2020] [Revised: 08/20/2020] [Accepted: 09/01/2020] [Indexed: 12/12/2022] Open
Abstract
Methylglyoxal (MG) is a dicarbonyl compound, the level of which is increased in the blood of diabetes patients. MG is reported to be involved in the development of cerebrovascular complications in diabetes, but the exact mechanisms need to be elucidated. Here, we investigated the possible roles of oxidative stress and mitophagy in MG-induced functional damage in brain endothelial cells (ECs). Treatment of MG significantly altered metabolic stress as observed by the oxygen-consumption rate and barrier-integrity as found in impaired trans-endothelial electrical resistance in brain ECs. The accumulation of MG adducts and the disturbance of the glyoxalase system, which are major detoxification enzymes of MG, occurred concurrently. Reactive oxygen species (ROS)-triggered oxidative damage was observed with increased mitochondrial ROS production and the suppressed Akt/hypoxia-inducible factor 1 alpha (HIF-1α) pathway. Along with the disturbance of mitochondrial bioenergetic function, parkin-1-mediated mitophagy was increased by MG. Treatment of N-acetyl cysteine significantly reversed mitochondrial damage and mitophagy. Notably, MG induced dysregulation of tight junction proteins including occludin, claudin-5, and zonula occluden-1 in brain ECs. Here, we propose that diabetic metabolite MG-associated oxidative stress may contribute to mitochondrial damage and autophagy in brain ECs, resulting in the dysregulation of tight junction proteins and the impairment of permeability.
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33
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Tang Y, Hu C, Sang S. Characterization of Reaction Products and Mechanisms between Serotonin and Methylglyoxal in Model Reactions and Mice. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 2020; 68:2437-2444. [PMID: 32011875 DOI: 10.1021/acs.jafc.0c00071] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/10/2023]
Abstract
Serotonin is an important endogenous regulatory neurotransmitter and has also been found in fruits, vegetables, and nuts. Methylglyoxal (MGO) is a reactive dicarbonyl metabolite and also a food toxin that modifies protein and DNA to cause the development of many chronic diseases. The objective of this study is to understand the reaction mechanisms between serotonin and MGO and determine whether serotonin could trap MGO in vivo. Five products were detected in phosphate buffer (pH 7.4) at 37 °C. Four products (compounds 2 and 4-6) were purified from the reaction mixture, and their structures were characterized by the analysis of their high-resolution mass and one- and two-dimensional nuclear magnetic resonance spectra. One product (compound 3), as a result of its instability, could not be properly purified and was tentatively characterized on the basis of its high-resolution mass spectrum and corresponding mass fragments. On the basis of the structures of these five products, two reaction pathways were proposed. Compounds 2, 3, 5, and 6 were produced through the Pictet-Spengler condensation pathway between the primary amine of serotonin and the ketone of MGO, and compound 3 was identified as the intermediate product to form products 2, 5, and 6, whereas compound 4 was formed through nucleophilic substitution by the benzene ring of serotonin, which is a new reaction pathway between biogenic amines and reactive carbonyl species. More importantly, the detection of adducts 2 and 4-6 in mice supports our hypothesis that the reaction between serotonin and MGO also happens in vivo through the same pathways as those in model reactions, suggesting that dietary or endogenous serotonin has the capacity to trap MGO in vivo.
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Affiliation(s)
- Yao Tang
- Laboratory for Functional Foods and Human Health, Center for Excellence in Post-Harvest Technologies , North Carolina Agricultural and Technical State University , North Carolina Research Campus, 500 Laureate Way , Kannapolis , North Carolina 28081 , United States
| | - Changling Hu
- Laboratory for Functional Foods and Human Health, Center for Excellence in Post-Harvest Technologies , North Carolina Agricultural and Technical State University , North Carolina Research Campus, 500 Laureate Way , Kannapolis , North Carolina 28081 , United States
| | - Shengmin Sang
- Laboratory for Functional Foods and Human Health, Center for Excellence in Post-Harvest Technologies , North Carolina Agricultural and Technical State University , North Carolina Research Campus, 500 Laureate Way , Kannapolis , North Carolina 28081 , United States
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Schalkwijk CG, Stehouwer CDA. Methylglyoxal, a Highly Reactive Dicarbonyl Compound, in Diabetes, Its Vascular Complications, and Other Age-Related Diseases. Physiol Rev 2020; 100:407-461. [DOI: 10.1152/physrev.00001.2019] [Citation(s) in RCA: 176] [Impact Index Per Article: 35.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023] Open
Abstract
The formation and accumulation of methylglyoxal (MGO), a highly reactive dicarbonyl compound, has been implicated in the pathogenesis of type 2 diabetes, vascular complications of diabetes, and several other age-related chronic inflammatory diseases such as cardiovascular disease, cancer, and disorders of the central nervous system. MGO is mainly formed as a byproduct of glycolysis and, under physiological circumstances, detoxified by the glyoxalase system. MGO is the major precursor of nonenzymatic glycation of proteins and DNA, subsequently leading to the formation of advanced glycation end products (AGEs). MGO and MGO-derived AGEs can impact on organs and tissues affecting their functions and structure. In this review we summarize the formation of MGO, the detoxification of MGO by the glyoxalase system, and the biochemical pathways through which MGO is linked to the development of diabetes, vascular complications of diabetes, and other age-related diseases. Although interventions to treat MGO-associated complications are not yet available in the clinical setting, several strategies to lower MGO have been developed over the years. We will summarize several new directions to target MGO stress including glyoxalase inducers and MGO scavengers. Targeting MGO burden may provide new therapeutic applications to mitigate diseases in which MGO plays a crucial role.
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Affiliation(s)
- C. G. Schalkwijk
- CARIM School for Cardiovascular Diseases, Maastricht University Medical Centre, Maastricht, The Netherlands; and Department of Internal Medicine, Maastricht University Medical Centre, Maastricht, The Netherlands
| | - C. D. A. Stehouwer
- CARIM School for Cardiovascular Diseases, Maastricht University Medical Centre, Maastricht, The Netherlands; and Department of Internal Medicine, Maastricht University Medical Centre, Maastricht, The Netherlands
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35
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Gong Y, Soleymani Abyaneh H, Drossis N, Niederquell A, Kuentz M, Leroux JC, de Haan HW, Gauthier MA. Ultra-sub-stoichiometric "Dynamic" Bioconjugation Reduces Viscosity by Disrupting Immunoglobulin Oligomerization. Biomacromolecules 2019; 20:3557-3565. [PMID: 31398010 DOI: 10.1021/acs.biomac.9b00867] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/17/2023]
Abstract
Monoclonal antibodies (mAb) are a major focus of the pharmaceutical industry, and polyclonal immunoglobulin G (IgG) therapy is used to treat a wide variety of health conditions. As some individuals require mAb/IgG therapy their entire life, there is currently a great desire to formulate antibodies for bolus injection rather than infusion. However, to achieve the required doses, very concentrated antibody solutions may be required. Unfortunately, mAb/IgG self-assembly at high concentration can produce an unacceptably high viscosity for injection. To address this challenge, this study expands the concept of "dynamic covalent chemistry" to "dynamic bioconjugation" in order to reduce viscosity by interfering with antibody-antibody interactions. Ultra-sub-stoichiometric amounts of dynamic PEGylation agents (down to the nanomolar) significantly reduced the viscosity of concentrated antibody solutions by interfering with oligomerization.
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Affiliation(s)
- Yuhui Gong
- Swiss Federal Institute of Technology Zurich (ETHZ) , Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences , Vladimir-Prelog-Weg 3 , 8093 Zurich , Switzerland
| | - Hoda Soleymani Abyaneh
- Institut National de la Recherche Scientifique (INRS) , EMT Research Center , 1650 boul. Lionel-Boulet , Varennes , J3X 1S2 , Canada
| | - Nicole Drossis
- University of Ontario Institute of Technology , Faculty of Science , Oshawa , Ontario L1H 7K4 , Canada
| | - Andreas Niederquell
- University of Applied Sciences Northwestern Switzerland , School of Life Sciences, Institute of Pharma Technology , Hofackerstr. 30 , 4132 Muttenz , Switzerland
| | - Martin Kuentz
- University of Applied Sciences Northwestern Switzerland , School of Life Sciences, Institute of Pharma Technology , Hofackerstr. 30 , 4132 Muttenz , Switzerland
| | - Jean-Christophe Leroux
- Swiss Federal Institute of Technology Zurich (ETHZ) , Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences , Vladimir-Prelog-Weg 3 , 8093 Zurich , Switzerland
| | - Hendrick W de Haan
- University of Ontario Institute of Technology , Faculty of Science , Oshawa , Ontario L1H 7K4 , Canada
| | - Marc A Gauthier
- Institut National de la Recherche Scientifique (INRS) , EMT Research Center , 1650 boul. Lionel-Boulet , Varennes , J3X 1S2 , Canada
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36
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Han L, Lin Q, Liu G, Han D, Niu L, Su D. Lipids Promote Glycated Phospholipid Formation by Inducing Hydroxyl Radicals in a Maillard Reaction Model System. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 2019; 67:7961-7967. [PMID: 31260294 DOI: 10.1021/acs.jafc.9b02771] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/09/2023]
Abstract
Food-derived glycated phospholipids is potentially hazardous to human health. However, there are few studies on the effects of lipids on the formation of glycated phospholipids. In this work, two model systems were established: (1) a model system including 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (PE), glucose, and Fenton reagent and (2) a model system including PE, glucose, and five kind of vegetable oils. The contents of carboxymethyl-PE, carboxyethyl-PE, Amadori-PE, hydroxyl radical (OH•), glyoxal, and methylglyoxal were determined with high-performance liquid chromatography mass spectrometry. The results of the first model system showed that OH• oxidized glucose to produce glyoxal and methylglyoxal, which then reacted with PE to form carboxymethyl-PE and carboxyethyl-PE. OH• also oxidized Amadori-PE to form carboxymethyl-PE. The results of the second model system showed that vegetable oils with higher number of moles of carbon-carbon unsaturated double bond in vegetable oil per kilogram could produce more OH•, which promote the formation of carboxymethyl-PE and carboxyethyl-PE by oxidizing glucose and oil. We elucidated the effects of oils on the formation of glycated phospholipids in terms of OH• and intermediates. This work will contribute to better understanding the formation mechanism of glycated phospholipids with oil.
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Affiliation(s)
- Lipeng Han
- Center for Advanced Analytical Science, School of Chemistry and Chemical Engineering , Guangzhou University , Guangzhou 510006 , China
- Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety , Guangzhou 510640 , China
| | - Qingna Lin
- School of Food Science and Engineering , South China University of Technology , Guangzhou 510640 , China
- Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety , Guangzhou 510640 , China
| | - Guoqin Liu
- School of Food Science and Engineering , South China University of Technology , Guangzhou 510640 , China
- Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety , Guangzhou 510640 , China
| | - Dongxue Han
- Center for Advanced Analytical Science, School of Chemistry and Chemical Engineering , Guangzhou University , Guangzhou 510006 , China
| | - Li Niu
- Center for Advanced Analytical Science, School of Chemistry and Chemical Engineering , Guangzhou University , Guangzhou 510006 , China
| | - Dongxiao Su
- Center for Advanced Analytical Science, School of Chemistry and Chemical Engineering , Guangzhou University , Guangzhou 510006 , China
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37
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Methylglyoxal – An emerging biomarker for diabetes mellitus diagnosis and its detection methods. Biosens Bioelectron 2019; 133:107-124. [DOI: 10.1016/j.bios.2019.03.010] [Citation(s) in RCA: 33] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/06/2018] [Revised: 03/07/2019] [Accepted: 03/07/2019] [Indexed: 02/07/2023]
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38
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Chen HJC, Teng YC. Stability of glyoxal- and methylglyoxal-modified hemoglobin on dried blood spot cards as analyzed by nanoflow liquid chromatography tandem mass spectrometry. J Food Drug Anal 2019; 27:526-530. [PMID: 30987724 PMCID: PMC9296192 DOI: 10.1016/j.jfda.2018.10.003] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/02/2018] [Revised: 10/04/2018] [Accepted: 10/05/2018] [Indexed: 12/17/2022] Open
Abstract
Blood sampling by the dried blood spot (DBS) technique has become commonly applied in newborn screening. It is often used for analysis of small molecules, such as metabolites. Recently, DBS sampling has been applied for quantification of post-translational protein modifications. Glyoxal and methylglyoxal are two simple oxoaldehydes released from glycated proteins in the Maillard reaction. They are widely distributed in the environment (e.g. cigarette smoke) and found in foods and beverages. Glyoxal and methylglyoxal are shown to react with biomolecules including DNA and proteins. In this laboratory, we previously identified the sites of modification by these two oxoaldehydes in human hemoglobin and found that the extents of modification at certain sites of lysine and arginine residues are significantly higher in type 2 diabetes mellitus patients than in nondiabetic individuals. In this study, we examine the stability of these modifications of hemoglobin stored on DBS cards at room temperature or 4 °C in the ambient air. After hemoglobin was extracted from the DBS cards, it was digested by trypsin and analyzed by nanoflow liquid chromatography coupled with nanospray ionization tandem mass spectrometry. The results show that the extents of all these PTMs are stable within 14 and 21 days when stored on DBS at room temperature and at 4 °C, respectively. Extraction of globin from DBS cards is mostly advantageous for hemolytic blood samples. This assay is sensitive as only a quarter of a DBS card containing ca. 12 μL of blood is required. Thus, it is practically useful to measure the extents of glyoxal- and methylglyoxal-induced hemoglobin modifications from DBS cards.
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Dhananjayan K, Irrgang F, Raju R, Harman DG, Moran C, Srikanth V, Münch G. Determination of glyoxal and methylglyoxal in serum by UHPLC coupled with fluorescence detection. Anal Biochem 2019; 573:51-66. [PMID: 30796906 DOI: 10.1016/j.ab.2019.02.014] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/06/2018] [Accepted: 02/16/2019] [Indexed: 01/17/2023]
Abstract
Glyoxal (GO) and methylglyoxal (MGO) are two important biomarkers in diabetes. Analytical methods for determination of GO and MGO in serum samples are either HPLC with UV-Vis (low sensitivity) or MS/MS (expensive) detection. These disadvantages have hampered the introduction of these biomarkers as a routine analyte for diabetes diagnostics into the clinical laboratory. In this study, we introduce a UHPLC method with fluorescence detection for the measurement of GO and MGO in serum samples by pre-column derivatization at neutral pH with 5, 6-diamino-2,4-dihydroxypyrimidine sulfate (DDP) to form lumazines. The method was validated as per FDA guidelines. Using this method, we have determined GO and MGO in a variety of animal serum samples, and for example, determined the GO and MGO concentration in adult bovine serum to be 852 ± 27 and 192 ± 10 nmol/L, respectively. In human serum, GO and MGO levels in non-diabetic subjects (n = 14) were determined to be 154 ± 88 and 98 ± 27 nmol/L, and in serum samples from subjects with diabetes (n = 14) 244 ± 137 and 190 ± 68 nmol/L, respectively. In addition, interference studies showed that physiological serum components did not lead to an artificial increase in the levels of GO and MGO.
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Affiliation(s)
- Karthik Dhananjayan
- Department of Pharmacology, School of Medicine, Western Sydney University, Locked Bag 1797, Penrith, NSW, 2751, Australia
| | - Felix Irrgang
- Department of Pharmacology, School of Medicine, Western Sydney University, Locked Bag 1797, Penrith, NSW, 2751, Australia
| | - Ritesh Raju
- Department of Pharmacology, School of Medicine, Western Sydney University, Locked Bag 1797, Penrith, NSW, 2751, Australia
| | - David G Harman
- Department of Pharmacology, School of Medicine, Western Sydney University, Locked Bag 1797, Penrith, NSW, 2751, Australia
| | - Chris Moran
- Department of Medicine, Peninsula Health & Peninsula Clinical School, Monash University, Victoria, 3199, Australia
| | - Velandai Srikanth
- Department of Medicine, Peninsula Health & Peninsula Clinical School, Monash University, Victoria, 3199, Australia; Menzies Institute for Medical Research, University of Tasmania, Hobart, Tasmania 7000, Australia
| | - Gerald Münch
- Department of Pharmacology, School of Medicine, Western Sydney University, Locked Bag 1797, Penrith, NSW, 2751, Australia; NICM Health Research Institute, Western Sydney University, Locked Bag 1797, Penrith, NSW, 2751, Australia.
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40
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Suh KS, Chon S, Jung WW, Choi EM. Crocin attenuates methylglyoxal-induced osteoclast dysfunction by regulating glyoxalase, oxidative stress, and mitochondrial function. Food Chem Toxicol 2019; 124:367-373. [DOI: 10.1016/j.fct.2018.12.031] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/17/2018] [Revised: 11/30/2018] [Accepted: 12/19/2018] [Indexed: 01/04/2023]
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41
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Tseng YT, Chang WH, Lin CC, Chang FR, Wu PC, Lo YC. Protective effects of Liuwei dihuang water extracts on diabetic muscle atrophy. PHYTOMEDICINE : INTERNATIONAL JOURNAL OF PHYTOTHERAPY AND PHYTOPHARMACOLOGY 2019; 53:96-106. [PMID: 30668418 DOI: 10.1016/j.phymed.2018.09.032] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 08/07/2018] [Revised: 08/20/2018] [Accepted: 09/03/2018] [Indexed: 06/09/2023]
Abstract
BACKGROUND Liuwei dihuang (LWDH), a widely used traditional Chinese herbal medicine, has been noticed for its potential on the improvement of diabetic complications including diabetic nephropathy and diabetic encephalopathy. However, whether LWDH can improve the effects of diabetic skeletal muscle atrophy has not yet been reported. PURPOSE The present study aimed to investigate the protective effects and mechanisms of the water extract of Liuwei dihuang (LWDH-WE) on skeletal muscle in cellular and animal models of diabetic muscle atrophy. STUDY DESIGN The muscle protective effects of LWDH-WE on diabetic muscle atrophy and weakness were examined in methylglyoxal (MG)-treated C2C12 myotubes and streptozotocin (STZ)-treated C57BL/6 mice, respectively. METHODS C2C12 myoblasts were differentiated by differentiation medium to form myotube structures. C2C12 myotubes were pre-treated with LWDH-WE 1 h before MG treatment. Diabetic mice were induced by single intraperitoneal injection of STZ (150 mg/kg) in C57BL/6 mice. Cell viability was determined by MTT and LDH assays. Protein expressions were detected by western blots. Morphological changes of cells were observed by an inverted microscope. Mitochondria membrane potential and reactive oxygen species (ROS) production were measured by flow cytometry. Muscle strength was evaluated by measuring grip strength of mice. RESULTS In C2C12 myotubes, LWDH-WE attenuated MG-induced cellular death and oxidative damage accompanied with improving mitochondrial membrane potential, inhibiting NADPH oxidase (Nox) activation, and ROS production. Moreover, LWDH-WE could attenuate MG-induced atrophy of C2C12 myotubes accompanied with regulating protein synthesis (IGF-1R, Akt, mTOR) and protein degradation (FoxO3a, atrogin-1, MuRF-1) signals. In STZ-induced diabetic mice, LWDH-WE improved body weight and skeletal muscle mass of mice. LWDH-WE also enhanced muscle strength of STZ-induced diabetic mice. Furthermore, LWDH-WE enhanced the improvement of insulin on gastrocnemius muscle mass and grip strength in STZ-treated mice. CONCLUSION LWDH-WE possesses skeletal muscle protection via reducing oxidative damage and regulating protein synthesis and degradation pathways in MG-induced atrophy of C2C12 myotubes. We also reveal the novel protection of LWDH-WE against STZ-induced reduction of muscle mass and muscle strength in mice.
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Affiliation(s)
- Yu-Ting Tseng
- Department of Pharmacology, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
| | - Wan-Hsuan Chang
- Department of Pharmacology, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
| | - Chih-Cheng Lin
- Department of Pharmacology, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
| | - Fang-Rong Chang
- Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
| | - Pao-Chu Wu
- School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan
| | - Yi-Ching Lo
- Department of Pharmacology, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
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Wang XJ, Ma SB, Liu ZF, Li H, Gao WY. Elevated levels of α-dicarbonyl compounds in the plasma of type II diabetics and their relevance with diabetic nephropathy. J Chromatogr B Analyt Technol Biomed Life Sci 2018; 1106-1107:19-25. [PMID: 30639946 DOI: 10.1016/j.jchromb.2018.12.027] [Citation(s) in RCA: 21] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/11/2018] [Revised: 12/19/2018] [Accepted: 12/27/2018] [Indexed: 02/02/2023]
Abstract
The presence of α‑dicarbonyl compounds (α-DCs) in vivo has been associated with the development of complications of diabetes mellitus (DM) and also with other chronic diseases. Therefore, quantitative analysis of α-DCs in body fluids is crucial to understand their roles in the formation of these chronic diseases. We established in this study a practical HPLC-UV method to measure 3‑deoxyglucosone (3-DG), glyoxal (GO), methylglyoxal (MGO), diacetyl (DA), and pentane‑2,3‑dione (PD) in blood plasma using 4‑(2,3‑dimethyl‑6‑quinoxalinyl)‑1,2‑benzenediamine (DQB) as a derivatizing reagent. The derivatizing reaction could be carried out quickly under mild conditions and the HPLC determination is simple, sensitive, and easy to operate. The recoveries of the α-DCs are between 85.26% and 110.20% (intra-day) and 87.25% and 103.18% (inter-day); the RSDs are between 1.28% and 5.69% (intra-day) and 2.26% and 6.34% (inter-day). We found the plasma levels of 3-DG, GO, and MGO in the diabetic patients are all significantly higher than those in healthy subjects. The results also show that the contents of GO and MGO in diabetic nephropathy (DN) patients are both significantly higher than those in simple T2DM patients. Moreover, it is found for the first time that the plasma level of GO might be a potential predictor of DN. The developed method would be useful for the measurements of the plasma α-DCs and the data acquired could be informative in the diagnosis of DM complications.
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Affiliation(s)
- Xin-Jie Wang
- National Engineering Research Center for Miniaturized Detection Systems and College of Life Sciences, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi 710069, China
| | - Sheng-Bo Ma
- National Engineering Research Center for Miniaturized Detection Systems and College of Life Sciences, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi 710069, China
| | - Zhuo-Feng Liu
- Friendship Hospital of Shaanxi Province, 227 West Youyi Road, Xi'an, Shaanxi 710000, China
| | - Heng Li
- National Engineering Research Center for Miniaturized Detection Systems and College of Life Sciences, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi 710069, China
| | - Wen-Yun Gao
- National Engineering Research Center for Miniaturized Detection Systems and College of Life Sciences, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi 710069, China.
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Chen HJC, Liu CT, Li YJ. Correlation between Glyoxal-Induced DNA Cross-Links and Hemoglobin Modifications in Human Blood Measured by Mass Spectrometry. Chem Res Toxicol 2018; 32:179-189. [PMID: 30507124 DOI: 10.1021/acs.chemrestox.8b00264] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/27/2022]
Abstract
Glyoxal is an oxoaldehyde generated from the degradation of glucose-protein conjugates and from lipid peroxidation in foods and in vivo, and it is also present in the environment (e.g., cigarette smoke). The major endogenous source of glyoxal is glucose autoxidation, and the glyoxal concentrations in plasma are higher in diabetic patients than in nondiabetics. Glyoxal reacts with biomolecules forming covalently modified DNA and protein adducts. We previously developed sensitive and specific assays based on nanoflow liquid chromatography-nanospray ionization tandem mass spectrometry (nanoLC-NSI/MS/MS) for quantification of DNA cross-linked adducts (dG-gx-dC and dG-gx-dA) and for hemoglobin adducts derived from glyoxal. In this study, we isolated and analyzed both leukocyte DNA and hemoglobin from the blood of diabetic patients and compared the adduct levels with those from nondiabetic subjects using the modified assays. The results indicated that the extents of glyoxal-induced hemoglobin modifications on α-Lys-11, α-Arg-92, β-Lys-17, and β-Lys-66 were statistically higher in diabetic patients than nondiabetics and they correlated with HbA1c significantly. Moreover, the levels of dG-gx-dC in leukocyte DNA correlated positively with the extents of globin modification at α-Lys-11 and β-Lys-17, while levels of dG-gx-dA correlated with those at α-Lys-11 and α-Arg-92 in nonsmoking subjects. Comparing the levels and the correlation coefficients of these hemoglobin and DNA adducts including or excluding smokers, it appears that smoking is not a major contributor to glyoxal-induced adduction of hemoglobin and leukocyte DNA. To the best of our knowledge, this is one of the few reports of positive correlation between DNA and protein adducts of the same compound (glyoxal) in the blood from the same subjects. Because of the high abundance of hemoglobin in blood, the results indicate that quantification of glyoxal-modified peptides in hemoglobin might serve as a dosimetry for glyoxal and a practical surrogate biomarker for assessing glyoxal-induced DNA damage and its prevention.
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Affiliation(s)
- Hauh-Jyun Candy Chen
- Department of Chemistry and Biochemistry , National Chung Cheng University , 168 University Road , Ming-Hsiung, Chia-Yi 62142 , Taiwan
| | - Chun-Ting Liu
- Department of Chemistry and Biochemistry , National Chung Cheng University , 168 University Road , Ming-Hsiung, Chia-Yi 62142 , Taiwan
| | - Yi-Jou Li
- Department of Chemistry and Biochemistry , National Chung Cheng University , 168 University Road , Ming-Hsiung, Chia-Yi 62142 , Taiwan
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Suh KS, Chon S, Jung WW, Choi EM. Effects of methylglyoxal on RANKL-induced osteoclast differentiation in RAW264.7 cells. Chem Biol Interact 2018; 296:18-25. [PMID: 30217477 DOI: 10.1016/j.cbi.2018.09.005] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/24/2018] [Revised: 07/27/2018] [Accepted: 09/11/2018] [Indexed: 12/20/2022]
Abstract
Methylglyoxal (MG) is a reactive dicarbonyl compound produced by glycolytic processing, which has been identified as a precursor of advanced glycation end products. Elevated MG levels in patients with diabetes are believed to contribute to diabetic complications, including bone defects. The objective of this study was to evaluate the effect of MG on RANKL-induced osteoclast differentiation in RAW264.7 cells, a murine macrophage cell line. RAW264.7 cells were cultured in medium containing 50 ng/mL RANKL and different concentrations of MG. Tartrate-resistant acid phosphatase (TRAP) activity and osteoclast bone resorbing activity were assessed and changes in intracellular calcium concentration, mitochondrial mass, mitochondrial membrane potential, and glyoxalase I level were examined. In addition, real-time RT-PCR assay was used to analyse osteoclast-associated genes. MG markedly inhibited RANKL-induced TRAP activity. MG treatment resulted in a significant decrease in intracellular calcium concentration, mitochondrial mass, mitochondrial membrane potential, and glyoxalase I level during osteoclastogenesis. In addition, MG increased the formation of mitochondrial superoxide. Quantitative reverse transcriptase-polymerase chain reaction revealed increased expression of the TRAF6, GAB2, ERK1, c-Fos, NFATc1, CLCN7, and OSTM1 genes, decreased expression of TCIRG and carbonic anhydrase II, and unchanged expression of cathepsin K and MMP-9 upon MG treatment. MG had no effect on the bone resorbing activity of osteoclasts. Our findings indicate that MG inhibits TRAP and glyoxalase I activity and impairs mitochondrial function in osteoclasts. Further validation of the underlying pathway is necessary.
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Affiliation(s)
- Kwang Sik Suh
- Department of Endocrinology & Metabolism, School of Medicine, Kyung Hee University, 1, Hoegi-dong, Dongdaemun-gu, Seoul, 02447, Republic of Korea
| | - Suk Chon
- Department of Endocrinology & Metabolism, School of Medicine, Kyung Hee University, 1, Hoegi-dong, Dongdaemun-gu, Seoul, 02447, Republic of Korea
| | - Woon-Won Jung
- Department of Biomedical Laboratory Science, College of Health Sciences, Cheongju University, Cheongju, Chungbuk, 28503, Republic of Korea
| | - Eun Mi Choi
- Department of Endocrinology & Metabolism, School of Medicine, Kyung Hee University, 1, Hoegi-dong, Dongdaemun-gu, Seoul, 02447, Republic of Korea.
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Banarjee R, Sharma A, Bai S, Deshmukh A, Kulkarni M. Proteomic study of endothelial dysfunction induced by AGEs and its possible role in diabetic cardiovascular complications. J Proteomics 2018; 187:69-79. [DOI: 10.1016/j.jprot.2018.06.009] [Citation(s) in RCA: 23] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/27/2018] [Revised: 06/12/2018] [Accepted: 06/15/2018] [Indexed: 12/30/2022]
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Holik AK, Stöger V, Hölz K, Somoza MM, Somoza V. Impact of free N ε-carboxymethyllysine, its precursor glyoxal and AGE-modified BSA on serotonin release from human parietal cells in culture. Food Funct 2018; 9:3906-3915. [PMID: 29972203 PMCID: PMC6053975 DOI: 10.1039/c8fo01045e] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/25/2018] [Accepted: 06/12/2018] [Indexed: 01/01/2023]
Abstract
Advanced glycation end products (AGEs) are frequently encountered in a western diet, in addition to their formation in vivo. N-Epsilon-carboxymethyllysine (CML), one of the chemically diverse compounds formed in the reaction between reducing carbohydrates and amines, is often used as a marker of advanced glycation, and has been shown to stimulate serotonin release from cells representing the central (SH-SY5Y cells) and the peripheral (Caco-2 cells) serotonin system in vitro. Here, we investigated the effect of glyoxal, free CML, and protein-linked AGE-BSA on serotonin release from human gastric tumour cells, which originate from an adenocarcinoma of the stomach and have recently been shown to be capable of serotonin synthesis and release. Microarray experiments showed both CML and glyoxal to alter genes associated with serotonin receptors. Furthermore, treatment with glyoxal resulted in a small change in RAGE expression while CML did not alter its expression. On a functional level, treatment with 500 μM CML increased extracellular serotonin content by 341 ± 241%, while treatment with 1 mg mL-1 AGE-BSA led to a reduction by 49 ± 11% compared to non-treated cells. The CML-induced serotonin release was reduced by the HTR3 antagonist granisetron. Incubation with the RAGE antagonist FPS-ZM1 abolished the effect of AGE-BSA on serotonin release, while no impact on CML-induced serotonin release was observed. Furthermore, treatment with 5 mM CML stimulated proton secretion as a functional outcome measure, assessed using a pH sensitive dye. Taken together, these results indicate a likely HTR3-mediated, RAGE-independent effect of free CML on serotonin release and a RAGE-dependent mechanism for the protein linked AGE-BSA.
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Affiliation(s)
- Ann-Katrin Holik
- Department of Physiological Chemistry
, Faculty of Chemistry
, University of Vienna
,
Althanstraße 14
, 1090 Vienna
, Austria
.
; Fax: +43 1 4277 9706
; Tel: +43 1 4227 70601
| | - Verena Stöger
- Christian Doppler Laboratory for Bioactive Aroma Compounds
, Faculty of Chemistry
, University of Vienna
,
Althanstraße 14
, 1090 Vienna
, Austria
| | - Kathrin Hölz
- Department of Inorganic Chemistry
, Faculty of Chemistry
, University of Vienna
,
Althanstraße 14
, 1090 Vienna
, Austria
| | - Mark M. Somoza
- Department of Inorganic Chemistry
, Faculty of Chemistry
, University of Vienna
,
Althanstraße 14
, 1090 Vienna
, Austria
| | - Veronika Somoza
- Department of Physiological Chemistry
, Faculty of Chemistry
, University of Vienna
,
Althanstraße 14
, 1090 Vienna
, Austria
.
; Fax: +43 1 4277 9706
; Tel: +43 1 4227 70601
- Christian Doppler Laboratory for Bioactive Aroma Compounds
, Faculty of Chemistry
, University of Vienna
,
Althanstraße 14
, 1090 Vienna
, Austria
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Reichert O, Fleming T, Neufang G, Schmelz M, Genth H, Kaever V, Wenck H, Stäb F, Terstegen L, Kolbe L, Roggenkamp D. Impaired glyoxalase activity is associated with reduced expression of neurotrophic factors and pro-inflammatory processes in diabetic skin cells. Exp Dermatol 2018; 26:44-50. [PMID: 27306297 DOI: 10.1111/exd.13118] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 06/01/2016] [Indexed: 12/18/2022]
Abstract
Patients suffering from type II diabetes develop several skin manifestations including cutaneous infections, diabetic dermopathy, diabetic bullae and acanthosis nigricans. Diabetic micro- and macroangiopathy as well as diabetic neuropathy are believed to play a crucial role in the development of diabetic skin disorders. A reduced cutaneous nerve fibre density was reported in diabetic subjects, which subsequently leads to impaired sensory nerve functions. Using an innervated skin model, we investigated the impact of human diabetic dermal fibroblasts and keratinocytes on porcine sensory neurons. Diabetic skin cells showed a reduced capacity to induce neurite outgrowth due to a decreased support with neurotrophic factors, such as NGF. Furthermore, diabetic keratinocytes displayed insulin resistance and increased expression of pro-inflammatory cytokines demonstrating the persistent effect of diabetes mellitus on human skin cells. Dysregulations were related to a significantly reduced glyoxalase enzyme activity in diabetic keratinocytes as experimentally reduced glyoxalase activity mimicked the increase in pro-inflammatory cytokine expression and reduction in NGF. Our results demonstrate an impaired crosstalk of diabetic skin cells and sensory neurons favouring hypo-innervation. We suggest that reduced methylglyoxal detoxification contributes to an impaired neurocutaneous interaction in diabetic skin.
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Affiliation(s)
- Olga Reichert
- Research and Development, Beiersdorf AG, Hamburg, Germany
| | - Thomas Fleming
- Department of Medicine I and Clinical Chemistry, University Hospital Heidelberg, Heidelberg, Germany
| | - Gitta Neufang
- Research and Development, Beiersdorf AG, Hamburg, Germany
| | - Martin Schmelz
- Department of Anesthesiology Mannheim, Heidelberg University, Mannheim, Germany
| | - Harald Genth
- Institute of Toxicology, Hannover Medical School, Hannover, Germany
| | - Volkhard Kaever
- Institute of Pharmacology, Hannover Medical School, Hannover, Germany
| | - Horst Wenck
- Research and Development, Beiersdorf AG, Hamburg, Germany
| | - Franz Stäb
- Research and Development, Beiersdorf AG, Hamburg, Germany
| | - Lara Terstegen
- Research and Development, Beiersdorf AG, Hamburg, Germany
| | - Ludger Kolbe
- Research and Development, Beiersdorf AG, Hamburg, Germany
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Kim JH, Kim KA, Shin YJ, Kim H, Majid A, Bae ON. Methylglyoxal induced advanced glycation end products (AGE)/receptor for AGE (RAGE)-mediated angiogenic impairment in bone marrow-derived endothelial progenitor cells. JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH. PART A 2018; 81:266-277. [PMID: 29473788 DOI: 10.1080/15287394.2018.1440185] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/08/2023]
Abstract
Endothelial cells (ECs) maintain the structure and function of blood vessels and are readily exposed to exogenous and endogenous toxic substances in the circulatory system. Bone marrow-derived endothelial progenitor cells (EPCs) circulate in the blood and differentiate to EC, which are known to participate in angiogenesis and regeneration of injured vessels. Dysfunction in EPC contributes to cardiovascular complications in patients with diabetes, but the precise molecular mechanisms underlying diabetic EPC abnormalities are not completely understood. The aim of this study was to investigate the mechanisms underlying diabetic EPC dysfunction using methylglyoxal (MG), an endogenous toxic diabetic metabolite. Data demonstrated that MG decreased cell viability and protein expression of vascular endothelial growth factor receptor (VEGFR)-2 associated with functional impairment of tube formation in EPC. The generation of advanced glycation end (AGE) products was increased in EPC following exposure to MG. Blockage of receptor for AGE (RAGE) by FPS-ZM1, a specific antagonist for RAGE, significantly reversed the decrease of VEGFR-2 protein expression and angiogenic dysfunction in MG-incubated EPC. Taken together, data demonstrated that MG induced angiogenic impairment in EPC via alterations in the AGE/RAGE-VEGFR-2 pathway which may be utilized in the development of potential therapeutic and preventive targets for diabetic vascular complications.
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Affiliation(s)
- Jeong-Hyeon Kim
- a College of Pharmacy Institute of Pharmaceutical Science and Technology , Hanyang University , Ansan , Republic of Korea
| | - Kyeong-A Kim
- a College of Pharmacy Institute of Pharmaceutical Science and Technology , Hanyang University , Ansan , Republic of Korea
| | - Young-Jun Shin
- a College of Pharmacy Institute of Pharmaceutical Science and Technology , Hanyang University , Ansan , Republic of Korea
| | - Haram Kim
- a College of Pharmacy Institute of Pharmaceutical Science and Technology , Hanyang University , Ansan , Republic of Korea
| | - Arshad Majid
- b Sheffield Institute for Translational Neuroscience , University of Sheffield , Sheffield , England
| | - Ok-Nam Bae
- a College of Pharmacy Institute of Pharmaceutical Science and Technology , Hanyang University , Ansan , Republic of Korea
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Shuck SC, Wuenschell GE, Termini JS. Product Studies and Mechanistic Analysis of the Reaction of Methylglyoxal with Deoxyguanosine. Chem Res Toxicol 2018; 31:105-115. [DOI: 10.1021/acs.chemrestox.7b00274] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
Affiliation(s)
- Sarah C. Shuck
- Department of Molecular Medicine, City of Hope and Beckman Research Institute, Duarte, California 91010, United States
| | - Gerald E. Wuenschell
- Department of Molecular Medicine, City of Hope and Beckman Research Institute, Duarte, California 91010, United States
| | - John S. Termini
- Department of Molecular Medicine, City of Hope and Beckman Research Institute, Duarte, California 91010, United States
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Chen HJC, Chang YL, Teng YC, Hsiao CF, Lin TS. A Stable Isotope Dilution Nanoflow Liquid Chromatography Tandem Mass Spectrometry Assay for the Simultaneous Detection and Quantification of Glyoxal-Induced DNA Cross-Linked Adducts in Leukocytes from Diabetic Patients. Anal Chem 2017; 89:13082-13088. [PMID: 29172486 DOI: 10.1021/acs.analchem.6b04296] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/20/2023]
Abstract
Glyoxal (gx) is a bifunctional electrophile capable of cross-linking DNA. Although it is present in foods and from the environment, endogenous formation of glyoxal occurs through metabolism of carbohydrates and oxidation of lipids and nucleic acids. Plasma concentrations of glyoxal are elevated in in diabetes mellitus patients compared to nondiabetics. The most abundant 2'-deoxyribonucleoside adducts cross-linked by glyoxal are dG-gx-dC, dG-gx-dA, and dG-gx-dG. These DNA cross-links can be mutagenic by damaging the integrity of the DNA structure. Herein, we developed a highly sensitive and specific assay for the simultaneous detection and quantification of the dG-gx-dC and dG-gx-dA cross-links based on stable isotope dilution (SID) nanoflow liquid chromatography nanospray ionization tandem mass spectrometry (nanoLC-NSI/MS/MS) under the highly selected reaction monitoring mode and using a triple quadrupole mass spectrometer. The entire assay procedure involved addition of the stable isotope standards [15N5]dG-gx-dC and [15N5]dG-gx-dA as internal standards, enzyme hydrolysis to release the cross-links as nucleosides, enrichment by a reversed-phase solid-phase extraction column, and nanoLC-NSI/MS/MS analysis. The detection limit is 0.19 amol for dG-gx-dC and 0.89 amol for dG-gx-dA, which is 400 and 80 times more sensitive, respectively, than capillary LC-NSI/MS/MS assay of these adducts. The lower limit of quantification was 94 and 90 amol for dG-gx-dC and dG-gx-dA, respectively, which is equivalent to 0.056 and 0.065 adducts in 108 normal nucleotides in 50 μg of DNA. In type 2 diabetes mellitus (T2DM) patients (n = 38), the levels of dG-gx-dC and dG-gx-dA in leukocyte DNA were 1.94 ± 1.20 and 2.10 ± 1.77 in 108 normal nucleotides, respectively, which were significantly higher than those in nondiabetics (n = 39: 0.83 ± 0.92 and 1.05 ± 0.99 in 108 normal nucleotides, respectively). Excluding the factor of smoking, an exogenous source of glyoxal, levels of these two cross-linked adducts were found to be significantly higher in nonsmoking T2DM patients than in nonsmoking control subjects. Furthermore, the levels of dG-gx-dC and dG-gx-dA correlated with HbA1c with statistical significance. To our best knowledge, this is the first report of the identification and quantification of glyoxal-derived cross-linked DNA adducts in human leukocyte DNA and their association with T2DM. This SID nanoLC-NSI/MS/MS assay is highly sensitive and specific and it requires only 50 μg of leukocyte DNA isolated from 2-3 mL of blood to accurately quantify these two cross-linked adducts simultaneously. Our assay thus provides a useful biomarker for the evaluation of glyoxal-derived DNA damage.
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Affiliation(s)
- Hauh-Jyun Candy Chen
- Department of Chemistry and Biochemistry, National Chung Cheng University , 168 University Road, Ming-Hsiung, Chia-Yi 62102, Taiwan
| | - Ya-Lang Chang
- Department of Chemistry and Biochemistry, National Chung Cheng University , 168 University Road, Ming-Hsiung, Chia-Yi 62102, Taiwan
| | - Yi-Chun Teng
- Department of Chemistry and Biochemistry, National Chung Cheng University , 168 University Road, Ming-Hsiung, Chia-Yi 62102, Taiwan
| | - Chiung-Fong Hsiao
- Department of Chemistry and Biochemistry, National Chung Cheng University , 168 University Road, Ming-Hsiung, Chia-Yi 62102, Taiwan
| | - Tsai-Shiuan Lin
- Department of Chemistry and Biochemistry, National Chung Cheng University , 168 University Road, Ming-Hsiung, Chia-Yi 62102, Taiwan
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