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Kawaida MY, Kwon OS, Ahn A, Reiter AS, Tillquist NM, Noh SG, Lee JW, Moore TE, Reed SA. Effects of an astaxanthin-containing supplement on oxidative status in skeletal muscle and circulation during deconditioning and reconditioning periods in polo ponies. Physiol Rep 2025; 13:e70346. [PMID: 40285451 PMCID: PMC12032451 DOI: 10.14814/phy2.70346] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/27/2024] [Revised: 04/09/2025] [Accepted: 04/11/2025] [Indexed: 04/29/2025] Open
Abstract
This study investigated the effects of astaxanthin (ASTX) supplementation on oxidative status during a deconditioning-reconditioning cycle. Twelve polo ponies were assigned to no supplementation (CON) or an ASTX supplemented group, which received oral administration of a supplement containing 75 mg ASTX daily for 32 weeks. Polo ponies underwent a 16-week deconditioning period (DECON) followed by a 16-week reconditioning program (RECON). Submaximal exercise tests (SETs) were performed at the beginning of the study (Baseline), after DECON, and after RECON. Blood samples were collected at -30, 0, 15, 30, and 60 min relative to each SET for oxidative status analysis. Muscle samples were collected 2 weeks before (Pre-Ex) and 2 h after (Post-Ex) each SET for muscle oxidative status and gene expression analyses. Pre-Ex muscles were analyzed for high-resolution respirometry. Circulating glutathione peroxidase (GPX) activity was increased (p ≤ 0.02) and protein carbonylation was decreased in ASTX (p ≤ 0.05). Muscle oxidative status was affected by DECON and reconditioning (p ≤ 0.05). ASTX increased gene expression of PPARGC1A after reconditioning (p ≤ 0.05). Deconditioning reduced oxidative phosphorylation at complex I and II (p = 0.01). Thus, a deconditioning-reconditioning cycle had greater impacts on muscle oxidative capacity than ASTX supplementation.
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Affiliation(s)
- Mia Y. Kawaida
- Department of Animal ScienceUniversity of ConnecticutStorrsConnecticutUSA
- Present address:
Department of Applied Physiology and KinesiologyUniversity of FloridaGainesvilleUSA
| | - Oh. Sung Kwon
- Department of KinesiologyUniversity of ConnecticutStorrsConnecticutUSA
- Department of Orthopaedic Surgery & Center on AgingUniversity of Connecticut School of MedicineFarmingtonConnecticutUSA
| | - Ahram Ahn
- Department of KinesiologyUniversity of ConnecticutStorrsConnecticutUSA
| | - Amanda S. Reiter
- Department of Animal ScienceUniversity of ConnecticutStorrsConnecticutUSA
- Present address:
Department of Animal ScienceTarleton State UniversityStephenvilleUSA
| | | | - Sung Gi Noh
- Department of KinesiologyUniversity of ConnecticutStorrsConnecticutUSA
| | - Jung W. Lee
- Department of StatisticsUniversity of ConnecticutStorrsConnecticutUSA
| | - Timothy E. Moore
- Statistical Consulting Services, Center for Open Research Resources & EquipmentUniversity of ConnecticutStorrsConnecticutUSA
| | - Sarah A. Reed
- Department of Animal ScienceUniversity of ConnecticutStorrsConnecticutUSA
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Giercuszkiewicz-Hecold B, Kulka M, Czopowicz M, Wilczak J, Szarska E, Strzelec K, Grzeczka A, Graczyk S, Hryniszyn A, Mularczyk M, Marycz K, Cywińska A. The effect of long term astaxanthin supplementation on the antioxidant status of racing Arabian horses - preliminary study. Sci Rep 2024; 14:27991. [PMID: 39543175 PMCID: PMC11564757 DOI: 10.1038/s41598-024-77732-9] [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: 07/24/2024] [Accepted: 10/24/2024] [Indexed: 11/17/2024] Open
Abstract
Astaxanthin due to its strong antioxidant activity is believed to reduce oxidative stress and therefore is considered as feed additive in pathological conditions and also for the athletes. It is promoted by several equine web portals, however, data supporting that concept in horses is limited. Thus, the aim of this study was to evaluate the effect of astaxanthin supplementation on the parameters of oxidative status in 3 years old, racing Arabian horses during long term observation and the changes related to a single training session of high intensity. Six horses were supplemented with astaxanthin at a dose of 0.52-0.58 mg/kg BW and 7 received no supplementation. Astaxanthin supplementation resulted in the increase in total antioxidant status by 31.5%, accompanied by decreases in the amount of total thiobarbituric acid-reactive substances -TBARS and glutathione reductases - GR values by 34.5% and 45.4%, respectively, after 1 month and this effect persisted until the end of the observation. After individual training session the activities of glutathione peroxidases and GR were lower by 69% and 46%, respectively, and TBARS lower by 38% in supplemented horses. These results directly confirmed the beneficial effects of astaxanthin supplementation on the antioxidant status of race horses. Astaxanthin partially counterbalance the training-related oxidative stress, save the horse natural antioxidant defense, and shift the redox status towards a more reducing environment. At the same time, exercise-induced reactive oxygen species production at certain level was maintained and so that contributed to training progress.
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Affiliation(s)
| | - Marek Kulka
- Department of Pathology and Veterinary Diagnostics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, 02-776, Warsaw, Poland
| | - Michał Czopowicz
- Division of Veterinary Epidemiology and Economics, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, 02-776, Warsaw, Poland
| | - Jacek Wilczak
- Department of Physiology, Institute of Veterinary Medicine, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, 02-776, Warsaw, Poland
| | - Ewa Szarska
- Military Institute of Hygiene and Epidemiology, Kozielska 4, 01-001, Warsaw, Poland
| | - Katarzyna Strzelec
- Department of Horse Breeding and Use, University of Life Sciences in Lublin, Akademicka 13, 20-950, Lublin, Poland
| | - Arkadiusz Grzeczka
- Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus University in Torun, Lwowska 1, 87-100, Torun, Poland
| | - Szymon Graczyk
- Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus University in Torun, Lwowska 1, 87-100, Torun, Poland
| | - Adrian Hryniszyn
- Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus University in Torun, Lwowska 1, 87-100, Torun, Poland
| | - Malwina Mularczyk
- International Institute of Translational Medicine, Jesionowa 11, 55-114, Malin, Wisznia Mała, Poland
| | - Krzysztof Marycz
- International Institute of Translational Medicine, Jesionowa 11, 55-114, Malin, Wisznia Mała, Poland
- Department of Veterinary Medicine and Epidemiology, Veterinary Institute for Regenerative Cures, School of Veterinary Medicine, University of California, Davis, CA, 95516, USA
| | - Anna Cywińska
- Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus University in Torun, Lwowska 1, 87-100, Torun, Poland.
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Zhang X, Geng A, Cao D, Dugarjaviin M. Identification of mulberry leaf flavonoids and evaluating their protective effects on H 2O 2-induced oxidative damage in equine skeletal muscle satellite cells. Front Mol Biosci 2024; 11:1353387. [PMID: 38650596 PMCID: PMC11033687 DOI: 10.3389/fmolb.2024.1353387] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/10/2023] [Accepted: 03/04/2024] [Indexed: 04/25/2024] Open
Abstract
Introduction: Horses are susceptible to oxidative stress during strenuous endurance exercise, leading to muscle fatigue and damage. Mulberry leaf flavonoids (MLFs) possess significant antioxidant properties. However, the antioxidant efficacy of MLFs can be influenced by the extraction process, and their impact on H2O2-induced oxidative stress in equine skeletal muscle satellite cells (ESMCs) remains unexplored. Methods: Our study employed three extraction methods to obtain MLFs: ultrasound-assisted extraction (CEP), purification with AB-8 macroporous resin (RP), and n-butanol extraction (NB-EP). We assessed the protective effects of these MLFs on H2O2-induced oxidative stress in ESMCs and analyzed the MLF components using metabolomics. Results: The results revealed that pre-treatment with MLFs dose-dependently protected ESMCs against H2O2-induced oxidative stress. The most effective concentrations were 0.8 mg/mL of CEP, 0.6 mg/mL of RP, and 0.6 mg/mL of NB-EP, significantly enhancing EMSC viability (p < 0.05). These optimized MLF concentrations promoted the GSH-Px, SOD and T-AOC activities (p < 0.05), while reducing MDA production (p < 0.05) in H2O2-induced ESMCs. Furthermore, these MLFs enhanced the gene expression, including Nrf2 and its downstream regulatory genes (TrxR1, GPX1, GPX3, SOD1, and SOD2) (p < 0.05). In terms of mitochondrial function, ESMCs pre-treated with MLFs exhibited higher basal respiration, spare respiratory capacity, maximal respiration, ATP-linked respiration compared to H2O2-induced ESMCs (p < 0.05). Additionally, MLFs enhanced cellular basal glycolysis, glycolytic reserve, and maximal glycolytic capacity (p < 0.05). Metabolomics analysis results revealed significant differences in mulberrin, kaempferol 3-O-glucoside [X-Mal], neohesperidin, dihydrokaempferol, and isobavachalcone among the three extraction processes (p < 0.05). Discussion: Our study revealed that MLFs enhance antioxidant enzyme activity, alleviate oxidative damage in ESMCs through the activation of the Nrf2 pathway, and improve mitochondrial respiration and cell energy metabolism. Additionally, we identified five potential antioxidant flavonoid compounds, suggesting their potential incorporation into the equine diet as a strategy to alleviate exercise-induced oxidative stress.
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Affiliation(s)
| | | | | | - Manglai Dugarjaviin
- lnner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science and Technology, Inner Mongolia Agricultural University, Hohhot, China
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Beghelli D, Zallocco L, Angeloni C, Bistoni O, Ronci M, Cavallucci C, Mazzoni MR, Nuccitelli A, Catalano C, Hrelia S, Lucacchini A, Giusti L. Dietary Supplementation with Boswellia serrata, Verbascum thapsus, and Curcuma longa in Show Jumping Horses: Effects on Serum Proteome, Antioxidant Status, and Anti-Inflammatory Gene Expression. Life (Basel) 2023; 13:life13030750. [PMID: 36983904 PMCID: PMC10055707 DOI: 10.3390/life13030750] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/04/2023] [Revised: 02/27/2023] [Accepted: 03/07/2023] [Indexed: 03/12/2023] Open
Abstract
Intense exercise can cause inflammation and oxidative stress due to the production of reactive oxygen species. These pathophysiological processes are interdependent, and each one can induce the other, creating a vicious circle. A placebo-controlled blind study was carried out in show jumping horses (n. 16) to evaluate the effects of a commercial dietary supplement (Dolhorse® N.B.F. Lanes srl, Milan, Italy) containing Verbascum thapsus leaf powder (1.42%), Curcuma longa (14.280 mg/kg), and Boswellia serrata (Roxb ex Colebr) (14.280 mg/kg) extracts. Before and after 10 days of dietary supplementation, blood samples were collected to evaluate the protein levels, antioxidants, and inflammatory responses by proteomic analysis or real-time Reverse Transcriptase-Polymerase Chain Reaction (real-time RT-PCR). A total of 36 protein spots, connected to 29 proteins, were modulated by dietary supplementation, whereas real-time RT-PCR revealed a significant downregulation of proinflammatory cytokines (interleukin 1α (p < 0.05) and interleukin-6 (0.005), toll-like receptor 4 (p < 0.05), and IKBKB (p < 0.05) in supplemented sport horses. Immunoglobulin chains, gelsolin, plasminogen, vitamin D binding protein, apolipoprotein AIV, and filamin B were overexpressed, whereas haptoglobin, α-2-HS-glycoprotein, α2-macroglobulin, afamin, amine oxidase, 60S acidic ribosomal protein, and complement fragments 3, 4, and 7 were reduced. No effect was observed on the antioxidant defense systems. The present results suggest this phytotherapy may reinforce the innate immune responses, thus representing a valid adjuvant to alleviate inflammation, which is a pathophysiological process in sport horses.
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Affiliation(s)
- Daniela Beghelli
- School of Biosciences and Veterinary Medicine, University of Camerino, 62032 Camerino, Italy
- Correspondence: (D.B.); (L.G.); Tel.: +39-737-403201 (D.B.); +39-737-402916 (L.G.)
| | - Lorenzo Zallocco
- Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56126 Pisa, Italy
| | - Cristina Angeloni
- Department for Life Quality Studies, Alma Mater Studiorum, University of Bologna, 47921 Rimini, Italy
| | - Onelia Bistoni
- Rheumatology Unit, Department of Medicine, University of Perugia, 06126 Perugia, Italy
| | - Maurizio Ronci
- Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara, 66100 Chieti, Italy
| | | | | | - Anna Nuccitelli
- School of Biosciences and Veterinary Medicine, University of Camerino, 62032 Camerino, Italy
| | | | - Silvana Hrelia
- Department for Life Quality Studies, Alma Mater Studiorum, University of Bologna, 47921 Rimini, Italy
| | - Antonio Lucacchini
- Department of Clinical and Experimental Medicine, University of Pisa, 56126 Pisa, Italy
| | - Laura Giusti
- School of Pharmacy, University of Camerino, 62032 Camerino, Italy
- Correspondence: (D.B.); (L.G.); Tel.: +39-737-403201 (D.B.); +39-737-402916 (L.G.)
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El-Sherbiny HR, Abdelnaby EA, El-Shahat KH, Salem NY, Ramadan ES, Yehia SG, Fathi M. Coenzyme Q10 Supplementation enhances testicular volume and hemodynamics, reproductive hormones, sperm quality, and seminal antioxidant capacity in goat bucks under summer hot humid conditions. Vet Res Commun 2022; 46:1245-1257. [PMID: 36048337 DOI: 10.1007/s11259-022-09991-8] [Citation(s) in RCA: 11] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/12/2022] [Accepted: 08/26/2022] [Indexed: 10/14/2022]
Abstract
Oxidative stress (OS) is brought on by heat stress (HS), which weakens antioxidant defense and initiates OS. Since mitochondria are the primary source of reactive oxygen species (ROS), HS-mediated OS may be lessened by targeting mitochondria with particular antioxidants. The purpose of this study was to investigate the effect of oral coenzyme Q10 (CoQ10) supplementation on the reproductive performance of goat bucks under HS conditions. Ten mature bucks were randomly separated into two groups and housed in an environment with a high-temperature humidity index (THI: 88.3 to 94.8; summer season). The first group (n = 5) got the baseline diet while the second group (n = 5) received supplemental oral CoQ10 (3 mg/kg BW; CoQ10 group) daily for six weeks. Testicular blood flow parameters (TBF), testicular volume (TV) and echogenicity (TE), nitric oxide (NO), seminal alanine aminotransferase (ALT) and catalase (CAT) activities, total antioxidant capacity (TAC), malondialdehyde (MDA) content, and semen quality traits were all measured. The examinations started a week before (W-1), on the first supplementation day (W0), and weekly for eight consecutive weeks (W1-W8). There were marked (P < 0.05) increases in TBF (W3-W6) and TV, and a decrease in TE (W3-W5) in the CoQ10 group compared to the CON group. Similarly, testosterone (T) and NO levels (W3-W5) in the CoQ10 group were higher (P < 0.05) than those of the control group. The CoQ10 group demonstrated significant (P < 0.05) increases in seminal CAT (W4-W8) and TAC (W2-W6) activities and decreases in ALT (W4-W7) activity and MDA (W5-W8) concentration as compared to the control group. The CoQ10 group showed improvements (P < 0.05) at W3-W6 for sperm progressive motility, viability, and normal morphology and at W6-W8 for sperm concentration. In conclusion, oral CoQ10 supplementation improved testicular hemodynamics, testosterone production, semen quality, and antioxidant capacity in goat bucks during summer heat stress conditions.
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Affiliation(s)
- Hossam R El-Sherbiny
- Theriogenology Department, Faculty of Veterinary Medicine, Cairo University, Giza square, Giza, 12211, Egypt.
| | - Elshymaa A Abdelnaby
- Theriogenology Department, Faculty of Veterinary Medicine, Cairo University, Giza square, Giza, 12211, Egypt
| | - K H El-Shahat
- Theriogenology Department, Faculty of Veterinary Medicine, Cairo University, Giza square, Giza, 12211, Egypt
| | - Noha Y Salem
- Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt
| | - Eman S Ramadan
- Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt
| | - Shimaa G Yehia
- Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt
| | - Mohamed Fathi
- Theriogenology Department, Faculty of Veterinary Medicine, Cairo University, Giza square, Giza, 12211, Egypt
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Oxidative Stress in Dairy Cows: Insights into the Mechanistic Mode of Actions and Mitigating Strategies. Antioxidants (Basel) 2021; 10:antiox10121918. [PMID: 34943022 PMCID: PMC8750585 DOI: 10.3390/antiox10121918] [Citation(s) in RCA: 19] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/03/2021] [Revised: 11/23/2021] [Accepted: 11/24/2021] [Indexed: 12/12/2022] Open
Abstract
This review examines several molecular mechanisms underpinning oxidative stress in ruminants and their effects on blood and milk oxidative traits. We also investigate strategies to alleviate or repair oxidative damages by improving animal immune functions using novel feed additives. Microbial pathogenic cells, feeding management, and body condition score were some of the studied factors, inducing oxidative stress in ruminants. The predominance of Streptococcus spp. (24.22%), Acinetobacter spp. (21.37%), Romboutsia spp. (4.99%), Turicibacter spp., (2.64%), Stenotrophomonas spp. (2.33%), and Enterococcus spp. (1.86%) was found in the microbiome of mastitis cows with a decrease of d-mannose and increase of xanthine:guanine ratio when Streptococcus increased. Diversity of energy sources favoring the growth of Fusobacterium make it a keystone taxon contributing to metritis. Ruminal volatile fatty acids rose with high-concentrate diets that decreased the ruminal pH, causing a lysis of rumen microbes and release of endotoxins. Moreover, lipopolysaccharide (LPS) concentration, malondialdehyde (MDA), and superoxide dismutase (SOD) activities increased in high concentrate cows accompanied by a reduction of total antioxidant capacity (T-AOC), glutathione peroxidase (GPx), and catalase (CAT) activity. In addition, albumin and paraoxonase concentrations were inversely related to oxidative stress and contributed to the protection of low-density and high-density lipoproteins against lipid peroxidation, protein carbonyl, and lactoperoxidase. High concentrate diets increased the expression of MAPK pro-inflammatory genes and decreased the expression of antioxidant genes and proteins in mammary epithelial tissues. The expression levels of NrF2, NQO1, MT1E, UGT1A1, MGST3, and MT1A were downregulated, whereas NF-kB was upregulated with a high-grain or high concentrate diet. Amino-acids, vitamins, trace elements, and plant extracts have shown promising results through enhancing immune functions and repairing damaged cells exposed to oxidative stress. Further studies comparing the long-term effect of synthetic feed additives and natural plant additives on animal health and physiology remain to be investigated.
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Henry ML, Velez-Irizarry D, Pagan JD, Sordillo L, Gandy J, Valberg SJ. The Impact of N-Acetyl Cysteine and Coenzyme Q10 Supplementation on Skeletal Muscle Antioxidants and Proteome in Fit Thoroughbred Horses. Antioxidants (Basel) 2021; 10:antiox10111739. [PMID: 34829610 PMCID: PMC8615093 DOI: 10.3390/antiox10111739] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2021] [Revised: 10/26/2021] [Accepted: 10/28/2021] [Indexed: 12/22/2022] Open
Abstract
Horses have one of the highest skeletal muscle oxidative capacities amongst mammals, which, combined with a high glycolytic capacity, could perturb redox status during maximal exercise. We determined the effect of 30 d of oral coenzyme Q10 and N-acetyl-cysteine supplementation (NACQ) on muscle glutathione (GSH), cysteine, ROS, and coenzyme Q10 concentrations, and the muscle proteome, in seven maximally exercising Thoroughbred horses using a placebo and randomized cross-over design. Gluteal muscle biopsies were obtained the day before and 1 h after maximal exercise. Concentrations of GSH, cysteine, coenzyme Q10, and ROS were measured, and citrate synthase, glutathione peroxidase, and superoxide dismutase activities analyzed. GSH increased significantly 1 h post-exercise in the NACQ group (p = 0.022), whereas other antioxidant concentrations/activities were unchanged. TMT proteomic analysis revealed 40 differentially expressed proteins with NACQ out of 387 identified, including upregulation of 13 mitochondrial proteins (TCA cycle and NADPH production), 4 Z-disc proteins, and down regulation of 9 glycolytic proteins. NACQ supplementation significantly impacted muscle redox capacity after intense exercise by enhancing muscle glutathione concentrations and increasing expression of proteins involved in the uptake of glutathione into mitochondria and the NAPDH-associated reduction of oxidized glutathione, without any evident detrimental effects on performance.
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Affiliation(s)
- Marisa L. Henry
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA; (D.V.-I.); (L.S.); (J.G.); (S.J.V.)
- Correspondence:
| | - Deborah Velez-Irizarry
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA; (D.V.-I.); (L.S.); (J.G.); (S.J.V.)
| | - Joe D. Pagan
- Kentucky Equine Research, Versailles, KY 40383, USA;
| | - Lorraine Sordillo
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA; (D.V.-I.); (L.S.); (J.G.); (S.J.V.)
| | - Jeff Gandy
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA; (D.V.-I.); (L.S.); (J.G.); (S.J.V.)
| | - Stephanie J. Valberg
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA; (D.V.-I.); (L.S.); (J.G.); (S.J.V.)
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