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1
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Chen M, Song L, Zeng A. Harnessing nature's arsenal: Targeting the TGF-β/Smad Cascade with novel natural anti-fibrotic agents. Fitoterapia 2025; 181:106372. [PMID: 39778722 DOI: 10.1016/j.fitote.2024.106372] [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: 07/15/2024] [Revised: 12/24/2024] [Accepted: 12/26/2024] [Indexed: 01/11/2025]
Abstract
BACKGROUND Hepatic fibrosis is a wound healing response that leads to excessive deposition of extracellular matrix (ECM) due to sustained liver injury. Hepatic stellate cells (HSCs) are key players in ECM synthesis, with the TGF-β/Smad signaling pathway being central to their activation. Despite advances in understanding the pathogenesis of hepatic fibrosis, effective anti-fibrotic therapies are still lacking. METHODS This treatise conducts a comprehensive review of the literature on the hepatoprotective effects of natural products, including natural medicine compounds, herbal extracts, and polysaccharides. The focus is on their ability to modulate the TGF-β pathway, which is critical in the activation of HSCs and ECM synthesis in hepatic fibrosis. RESULTS The review identifies a variety of natural products that have shown promise in inhibiting the TGF-β/Smad signaling cascade, thereby reducing the activation of HSCs and ECM accumulation. These findings highlight the potential of these natural products as therapeutic agents in the treatment of hepatic fibrosis. CONCLUSIONS The exploration of natural products as modulators of the TGF-β pathway presents a novel avenue for both clinical and preclinical research into hepatic fibrosis. Further investigation is warranted to fully understand the mechanisms of action and to develop these compounds into effective anti-fibrotic pharmaceuticals.
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Affiliation(s)
- Maohua Chen
- Translational Chinese Medicine Key Laboratory of Sichuan Province, Sichuan Academy of Chinese Medicine Sciences, Sichuan Institute for Translational Chinese Medicine,Chengdu, Sichuan 610041, PR China; Department of Plastic Surgery, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China. Chengdu, Sichuan 610072, PR China
| | - Linjiang Song
- School of Medical and Life Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 611137, PR China.
| | - Anqi Zeng
- Translational Chinese Medicine Key Laboratory of Sichuan Province, Sichuan Academy of Chinese Medicine Sciences, Sichuan Institute for Translational Chinese Medicine,Chengdu, Sichuan 610041, PR China.
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2
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Ilha M, Meira Martins LA, da Silveira Moraes K, Dias CK, Thomé MP, Petry F, Rohden F, Borojevic R, Trindade VMT, Klamt F, Barbé‐Tuana F, Lenz G, Guma FCR. Caveolin-1 influences mitochondrial plasticity and function in hepatic stellate cell activation. Cell Biol Int 2022; 46:1787-1800. [PMID: 35971753 PMCID: PMC9804617 DOI: 10.1002/cbin.11876] [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: 01/15/2022] [Revised: 03/21/2022] [Accepted: 05/26/2022] [Indexed: 01/05/2023]
Abstract
Caveolin-1 (Cav-1) is an integral membrane protein present in all organelles, responsible for regulating and integrating multiple signals as a platform. Mitochondria are extremely adaptable to external cues in chronic liver diseases, and expression of Cav-1 may affect mitochondrial flexibility in hepatic stellate cells (HSCs) activation. We previously demonstrated that exogenous expression of Cav-1 was sufficient to increase some classical markers of activation in HSCs. Here, we aimed to evaluate the influence of exogenous expression and knockdown of Cav-1 on regulating the mitochondrial plasticity, metabolism, endoplasmic reticulum (ER)-mitochondria distance, and lysosomal activity in HSCs. To characterize the mitochondrial, lysosomal morphology, and ER-mitochondria distance, we perform transmission electron microscope analysis. We accessed mitochondria and lysosomal networks and functions through a confocal microscope and flow cytometry. The expression of mitochondrial machinery fusion/fission genes was examined by real-time polymerase chain reaction. Total and mitochondrial cholesterol content was measured using Amplex Red. To define energy metabolism, we used the Oroboros system in the cells. We report that GRX cells with exogenous expression or knockdown of Cav-1 changed mitochondrial morphometric parameters, OXPHOS metabolism, ER-mitochondria distance, lysosomal activity, and may change the activation state of HSC. This study highlights that Cav-1 may modulate mitochondrial function and structural reorganization in HSC activation, being a potential candidate marker for chronic liver diseases and a molecular target for therapeutic intervention.
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Affiliation(s)
- Mariana Ilha
- Programa de Pós‐Graduação em Ciências Biológicas‐Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto AlegreRio Grande do SulBrasil,Department of Clinical Nutrition, Institute of Public Health and Clinical NutritionUniversity of Eastern FinlandKuopioFinland
| | - Leo A. Meira Martins
- Programa de Pós‐Graduação em Ciências Biológicas‐Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto AlegreRio Grande do SulBrasil,Departamento de Fisiologia, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul ‐ UFRGSPorto AlegreRio Grande do SulBrasil
| | - Ketlen da Silveira Moraes
- Departamento de Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul ‐ UFRGSPorto AlegreRio Grande do SulBrasil
| | - Camila K. Dias
- Programa de Pós‐Graduação em Ciências Biológicas‐Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto AlegreRio Grande do SulBrasil
| | - Marcos P. Thomé
- Departamento de Biofísica e Centro de BiotecnologiaUniversidade Federal do Rio Grande do Sul ‐ UFRGSPorto AlegreRio Grande do SulBrasil
| | - Fernanda Petry
- Programa de Pós‐Graduação em Ciências Biológicas‐Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto AlegreRio Grande do SulBrasil
| | - Francieli Rohden
- Programa de Pós‐Graduação em Ciências Biológicas‐Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto AlegreRio Grande do SulBrasil
| | - Radovan Borojevic
- Centro de Medicina RegenerativaFaculdade Arthur Sa Earp Neto ‐ Faculdade de Medicina de PetrópolisRio de JaneiroBrasil
| | - Vera M. T. Trindade
- Departamento de Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul ‐ UFRGSPorto AlegreRio Grande do SulBrasil
| | - Fábio Klamt
- Departamento de Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul ‐ UFRGSPorto AlegreRio Grande do SulBrasil
| | - Florência Barbé‐Tuana
- Programa de Pós‐Graduação em Biologia Celular e MolecularEscola de Ciências da Pontifícia Universidade Católica do Rio Grande do Sul‐ PUCRSPorto AlegreRio Grande do SulBrasil
| | - Guido Lenz
- Departamento de Biofísica e Centro de BiotecnologiaUniversidade Federal do Rio Grande do Sul ‐ UFRGSPorto AlegreRio Grande do SulBrasil
| | - Fátima C. R. Guma
- Departamento de Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul ‐ UFRGSPorto AlegreRio Grande do SulBrasil,Centro de Microscopia e MicroanáliseUniversidade Federal do Rio Grande do Sul ‐ UFRGSPorto AlegreRio Grande do SulBrasil
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3
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Dai W, Qin Q, Li Z, Lin L, Li R, Fang Z, Han Y, Mu W, Ren L, Liu T, Zhan X, Xiao X, Bai Z. Curdione and Schisandrin C Synergistically Reverse Hepatic Fibrosis via Modulating the TGF-β Pathway and Inhibiting Oxidative Stress. Front Cell Dev Biol 2021; 9:763864. [PMID: 34858986 PMCID: PMC8631446 DOI: 10.3389/fcell.2021.763864] [Citation(s) in RCA: 14] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/24/2021] [Accepted: 10/12/2021] [Indexed: 11/22/2022] Open
Abstract
Hepatic fibrosis is the final pathway of several chronic liver diseases, which is characterized by the accumulation of extracellular matrix due to chronic hepatocyte damage. Activation of hepatic stellate cells and oxidative stress (OS) play an important role in mediating liver damage and initiating hepatic fibrosis. Hence, hepatic fibrosis can be reversed by inhibiting multiple channels such as oxidative stress, liver cell damage, or activation of hepatic stellate cells. Liuwei Wuling Tablets is a traditional Chinese medicine formula with the effect of anti- hepatic fibrosis, but the composition and mechanism of reversing hepatic fibrosis are still unclear. Our study demonstrated that one of the main active components of the Chinese medicine Schisandra chinensis, schisandrin C (Sin C), significantly inhibited oxidative stress and prevented hepatocyte injury. Meanwhile one of the main active components of the Chinese medicine Curdione inhibited hepatic stellate cell activation by targeting the TGF-β1/Smads signaling pathway. The further in vivo experiments showed that Sin C, Curdione and the combination of both have the effect of reversing liver fibrosis in mice, and the combined effect of inhibiting hepatic fibrosis is superior to treatment with Sin C or Curdione alone. Our study provides a potential candidate for multi-molecular or multi-pathway combination therapies for the treatment of hepatic fibrosis and demonstrates that combined pharmacotherapy holds great promise in the prevention and treatment of hepatic fibrosis.
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Affiliation(s)
- Wenzhang Dai
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China.,School of Pharmacy, Hunan University of Chinese Medicine, Changsha, China
| | - Qin Qin
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China
| | - Zhiyong Li
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China
| | - Li Lin
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China
| | - Ruisheng Li
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China
| | - Zhie Fang
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China
| | - Yanzhong Han
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China
| | - Wenqing Mu
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China
| | - Lutong Ren
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China
| | - Tingting Liu
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China
| | - Xiaoyan Zhan
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China.,China Military Institute of Chinese Materia, The Fifth Medical Centre, Chinese PLA General Hospital, Beijing, China
| | - Xiaohe Xiao
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China.,School of Pharmacy, Hunan University of Chinese Medicine, Changsha, China.,China Military Institute of Chinese Materia, The Fifth Medical Centre, Chinese PLA General Hospital, Beijing, China
| | - Zhaofang Bai
- Senior Department of Hepatology, The Fifth Medical Center of Chinese PLA General Hospital, Beijing, China.,China Military Institute of Chinese Materia, The Fifth Medical Centre, Chinese PLA General Hospital, Beijing, China
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4
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Gonçalves da Silva EF, Costa BP, Nassr MT, de Souza Basso B, Bastos MS, Antunes GL, Reghelin CK, Rosa Garcia MC, Schneider Levorse VG, Carlessi LP, Antunes Fernandes KH, Richter Schmitz CR, Haute GV, Luft C, Santarém E, Barbé-Tuana FM, Donadio MVF, Basso LA, Machado P, Rodrigues de Oliveira J. Therapeutic effect of uridine phosphorylase 1 (UPP1) inhibitor on liver fibrosis in vitro and in vivo. Eur J Pharmacol 2020; 890:173670. [PMID: 33098831 DOI: 10.1016/j.ejphar.2020.173670] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/18/2019] [Revised: 10/19/2020] [Accepted: 10/21/2020] [Indexed: 02/07/2023]
Abstract
Potassium 5-cyano-4-methyl-6-oxo-1,6-dihydropyridine-2-olate (CPBMF65) is a potent inhibitor of the uridine phosphorylase 1 (UPP1) enzyme. Its non-ionized analog has already demonstrated biological properties by reducing adverse effects caused by the chemotherapeutic 5-fluorouracil (5-FU). In addition, it has been demonstrated that uridine inhibits inflammation and fibrosis in bleomycin lung injury, decreasing collagen production. The purpose of this study was to investigate the in vitro and in vivo effects of CPBMF65 on activated hepatic stellate cells (HSC) and on carbon tetrachloride-induced liver fibrosis in mice. After incubation with CPBMF65, decreased cell proliferation and phenotype reversion were observed in vitro. In addition, CPBMF65 promoted a protective effect on tetrachloride-induced liver fibrosis in mice, demonstrated by its antifibrotic and anti-inflammatory actions. The results of the present study indicate that the UPP1 inhibitor (CPBMF65) may have potential as a novel therapeutic agent for the treatment of liver fibrosis.
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Affiliation(s)
- Elisa Feller Gonçalves da Silva
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil.
| | - Bruna Pasqualotto Costa
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Marcella Tornquist Nassr
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Bruno de Souza Basso
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Matheus Scherer Bastos
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Géssica Luana Antunes
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Camille Kirinus Reghelin
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Maria Claudia Rosa Garcia
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Vitor Giancarlo Schneider Levorse
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Leonardo Pfeiff Carlessi
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Krist Helen Antunes Fernandes
- Laboratório de Imunologia Clínica e Experimental, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Carine Raquel Richter Schmitz
- Programa de Pós-Graduação Em Biologia Celular: Bioquímica, Universidade Federal Do Rio Grande Do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Gabriela Viegas Haute
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Carolina Luft
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Eliane Santarém
- Laboratório de Biotecnologia Vegetal, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Florencia María Barbé-Tuana
- Laboratório de Imunobiologia, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Márcio Vinícius Fagundes Donadio
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Luiz Augusto Basso
- Centro de Pesquisas Em Biologia Molecular e Funcional (CPBMF), Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), TecnoPuc, Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Pablo Machado
- Centro de Pesquisas Em Biologia Molecular e Funcional (CPBMF), Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), TecnoPuc, Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
| | - Jarbas Rodrigues de Oliveira
- Laboratório de Pesquisa Em Biofísica Celular e Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, 90619-900, Brazil
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5
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de Oliveira CM, Martins LAM, de Sousa AC, Moraes KDS, Costa BP, Vieira MQ, Coelho BP, Borojevic R, de Oliveira JR, Guma FCR. Resveratrol increases the activation markers and changes the release of inflammatory cytokines of hepatic stellate cells. Mol Cell Biochem 2020; 476:649-661. [PMID: 33073314 DOI: 10.1007/s11010-020-03933-1] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/26/2020] [Accepted: 10/07/2020] [Indexed: 02/07/2023]
Abstract
The phytoalexin Resveratrol (3,5,4'-trihydroxystilbene; RSV) has been related to numerous beneficial effects on health by its cytoprotection and chemoprevention activities. Liver fibrosis is characterized by the extracellular matrix accumulation after hepatic injury and can lead to cirrhosis. Hepatic stellate cells (HSC) play a crucial role during fibrogenesis and liver wound healing by changing their quiescent phenotype to an activated phenotype for protecting healthy areas from damaged areas. Strategies on promoting the activated HSC death, the quiescence return or the cellular activation stimuli decrease play an important role on reducing liver fibrosis. Here, we evaluated the RSV effects on some markers of activation in GRX, an HSC model. We further evaluated the RSV influence in the ability of GRX on releasing inflammatory mediators. RSV at 1 and 10 µM did not alter the protein content of α-SMA, collagen I and GFAP; but 50 µM increased the content of these activation-related proteins. Also, RSV did not change the myofibroblast-like morphology of GRX. Interestingly, RSV at 10 and 50 µM decreased the GRX migration and collagen-I gel contraction. Finally, we showed that RSV triggered the increase in the TNF-α and IL-10 content in culture media of GRX while the opposite occurred for the IL-6 content. Altogether, these results suggested that RSV did not decrease the activation state of GRX and oppositely, triggered a pro-activation effect at the 50 µM concentration. However, despite the increase of TNF- α in culture media, these results on IL-6 and IL-10 secretion were in accordance with the anti-inflammatory role of RSV in our model.
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Affiliation(s)
- Cleverson Moraes de Oliveira
- Departmento de Bioquímica, ICBS, Universidade Federal Do Rio Grande Do Sul (UFRGS), Rua Ramiro Barcelos, 2600-Anexo I, Porto Alegre, RS, CEP, 90035-003, Brazil.
| | - Leo Anderson Meira Martins
- Departmento de Bioquímica, ICBS, Universidade Federal Do Rio Grande Do Sul (UFRGS), Rua Ramiro Barcelos, 2600-Anexo I, Porto Alegre, RS, CEP, 90035-003, Brazil.,Departamento de Fisiologia, ICBS, Universidade Federal Do Rio Grande Do Sul, Rua Sarmento Leite, Porto Alegre, RS, CEP, 500, Brazil
| | - Arieli Cruz de Sousa
- Departmento de Bioquímica, ICBS, Universidade Federal Do Rio Grande Do Sul (UFRGS), Rua Ramiro Barcelos, 2600-Anexo I, Porto Alegre, RS, CEP, 90035-003, Brazil
| | - Ketlen da Silveira Moraes
- Departmento de Bioquímica, ICBS, Universidade Federal Do Rio Grande Do Sul (UFRGS), Rua Ramiro Barcelos, 2600-Anexo I, Porto Alegre, RS, CEP, 90035-003, Brazil
| | - Bruna Pasqualotto Costa
- Laboratório de Pesquisa Em Biofísica Celular E Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, RS, Brazil
| | - Moema Queiroz Vieira
- Departmento de Bioquímica, ICBS, Universidade Federal Do Rio Grande Do Sul (UFRGS), Rua Ramiro Barcelos, 2600-Anexo I, Porto Alegre, RS, CEP, 90035-003, Brazil
| | - Bárbara Paranhos Coelho
- Departmento de Bioquímica, ICBS, Universidade Federal Do Rio Grande Do Sul (UFRGS), Rua Ramiro Barcelos, 2600-Anexo I, Porto Alegre, RS, CEP, 90035-003, Brazil
| | - Radovan Borojevic
- Centro de Medicina Regenerativa, Faculdade de Medicina de Petrópolis, Petrópolis, RJ, Brazil
| | - Jarbas Rodrigues de Oliveira
- Laboratório de Pesquisa Em Biofísica Celular E Inflamação, Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), Porto Alegre, RS, Brazil
| | - Fátima Costa Rodrigues Guma
- Departmento de Bioquímica, ICBS, Universidade Federal Do Rio Grande Do Sul (UFRGS), Rua Ramiro Barcelos, 2600-Anexo I, Porto Alegre, RS, CEP, 90035-003, Brazil.,Centro de Microscopia E Microanálise (CMM), Universidade Federal Do Rio Grande Do Sul, Av. Bento Gonçalves, 9500 - Prédio 43.177 - Bl 1Campus do Vale, Porto Alegre, RS, CEP, 91501-970, Brazil
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6
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Positive Effects of Ger-Gen-Chyn-Lian-Tang on Cholestatic Liver Fibrosis in Bile Duct Ligation-Challenged Mice. Int J Mol Sci 2019; 20:ijms20174181. [PMID: 31455001 PMCID: PMC6747316 DOI: 10.3390/ijms20174181] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/05/2019] [Revised: 08/12/2019] [Accepted: 08/24/2019] [Indexed: 01/22/2023] Open
Abstract
The purpose of this study was to investigate whether Ger-Gen-Chyn-Lian-Tang (GGCLT) suppresses oxidative stress, inflammation, and angiogenesis during experimental liver fibrosis through the hypoxia-inducible factor-1α (HIF-1α)-mediated pathway. Male C57BL/6 mice were randomly assigned to a sham-control or bile duct ligation (BDL) group with or without treatment with GGCLT at 30, 100, and 300 mg/kg. Plasma alanine aminotransferase (ALT) levels were analyzed using a diagnostic kit. Liver histopathology and hepatic status parameters were measured. Compared to control mice, the BDL mice exhibited an enlargement in liver HIF-1α levels, which was suppressed by 100 and 300 mg/kg GGCLT treatments (control: BDL: BDL + GGCLT-100: BDL + GGCLT-300 = 0.95 ± 0.07: 1.95 ± 0.12: 1.43 ± 0.05: 1.12 ± 0.10 fold; p < 0.05). GGCLT restrained the induction of hepatic hydroxyproline and malondialdehyde levels in the mice challenged with BDL, further increasing the hepatic glutathione levels. Furthermore, in response to increased hepatic inflammation and fibrogenesis, significant levels of ALT, nuclear factor kappa B, transforming growth factor-β, α-smooth muscle actin, matrix metalloproteinase-2 (MMP-2), MMP-9, and procollagen-III were found in BDL mice, which were attenuated with GGCLT. In addition, GGCLT reduced the induction of angiogenesis in the liver after BDL by inhibiting vascular endothelial growth factor (VEGF) and VEGF receptors 1 and 2. In conclusion, the anti-liver fibrosis effect of GGCLT, which suppresses hepatic oxidative stress and angiogenesis, may be dependent on an HIF-1α-mediated pathway.
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7
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Ilha M, Moraes KDS, Rohden F, Martins LAM, Borojevic R, Lenz G, Barbé‐Tuana F, Guma FCR. Exogenous expression of caveolin‐1 is sufficient for hepatic stellate cell activation. J Cell Biochem 2019; 120:19031-19043. [DOI: 10.1002/jcb.29226] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/12/2019] [Accepted: 06/04/2019] [Indexed: 12/18/2022]
Affiliation(s)
- Mariana Ilha
- Programa de Pós‐Graduação em Ciências Biológicas‐ Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto Alegre RS Brazil
| | - Ketlen da Silveira Moraes
- Programa de Pós‐Graduação em Ciências Biológicas‐ Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto Alegre RS Brazil
| | - Francieli Rohden
- Programa de Pós‐Graduação em Ciências Biológicas‐ Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto Alegre RS Brazil
| | - Leo Anderson Meira Martins
- Programa de Pós‐Graduação em Ciências Biológicas‐ Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto Alegre RS Brazil
| | - Radovan Borojevic
- Centro de Medicina RegenerativaFaculdade de Medicina de Petrópolis – FMPPetrópolis RJ Brazil
| | - Guido Lenz
- Departamento de Biofísica e Centro de BiotecnologiaUniversidade Federal do Rio Grande do Sul ‐ UFRGSPorto Alegre RS Brazil
| | - Florencia Barbé‐Tuana
- Programa de Pós‐Graduação em Ciências Biológicas‐ Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto Alegre RS Brazil
- Programa de Pós‐Graduação em Biologia Celular e MolecularEscola de Ciências da Pontifícia Universidade Católica do Rio Grande do Sul‐ PUCRSPorto Alegre RS Brazil
| | - Fátima Costa Rodrigues Guma
- Programa de Pós‐Graduação em Ciências Biológicas‐ Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do Sul – UFRGSPorto Alegre RS Brazil
- Centro de Microscopia e MicroanáliseUniversidade Federal do Rio Grande do Sul ‐ UFRGSPorto Alegre RS Brazil
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Colares JR, Schemitt EG, Hartmann RM, Licks F, Soares MDC, Bosco AD, Marroni NP. Antioxidant and anti-inflammatory action of melatonin in an experimental model of secondary biliary cirrhosis induced by bile duct ligation. World J Gastroenterol 2016; 22:8918-8928. [PMID: 27833383 PMCID: PMC5083797 DOI: 10.3748/wjg.v22.i40.8918] [Citation(s) in RCA: 36] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/28/2016] [Revised: 08/24/2016] [Accepted: 09/14/2016] [Indexed: 02/06/2023] Open
Abstract
AIM To evaluate the effects of melatonin (Mel) on oxidative stress in an experimental model of bile duct ligation (BDL). METHODS Male Wistar rats (n = 32, weight ± 300 g) were allocated across four groups: CO (sham BDL), BDL (BDL surgery), CO + Mel (sham BDL and Mel administration) and BDL + Mel (BDL surgery and Mel administration). Mel was administered intraperitoneally for 2 wk, starting on postoperative day 15, at a dose of 20 mg/kg. RESULTS Mel was effective at the different standards, reestablishing normal liver enzyme levels, reducing the hepatosomatic and splenosomatic indices, restoring lipoperoxidation and antioxidant enzyme concentrations, reducing fibrosis and inflammation, and thereby reducing liver tissue injury in the treated animals. CONCLUSION The results of this study suggest a protective effect of Mel when administered to rats with secondary biliary cirrhosis induced by BDL.
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de Souza ICC, Martins LAM, de Vasconcelos M, de Oliveira CM, Barbé-Tuana F, Andrade CB, Pettenuzzo LF, Borojevic R, Margis R, Guaragna R, Guma FCR. Resveratrol Regulates the Quiescence-Like Induction of Activated Stellate Cells by Modulating the PPARγ/SIRT1 Ratio. J Cell Biochem 2016; 116:2304-12. [PMID: 25833683 DOI: 10.1002/jcb.25181] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/07/2015] [Accepted: 03/31/2015] [Indexed: 01/27/2023]
Abstract
The activation of hepatic stellate cell (HSC), from a quiescent cell featuring cytoplasmic lipid droplets to a proliferative myofibroblast, plays an important role in liver fibrosis development. The GRX line is an activated HSC model that can be induced by all-trans-retinol to accumulate lipid droplets. Resveratrol is known for activating Sirtuin1 (SIRT1), a NAD(+)-dependent deacetylase that suppresses the activity of peroxisome proliferator-activated receptor gamma (PPARγ), an important adipogenic transcription factor involved in the quiescence maintenance of HSC. We evaluated the effects of 0.1 μM of resveratrol in retinol-induced GRX quiescence by investigating the interference of SIRT1 and PPARγ on cell lipogenesis. GRX lipid accumulation was evaluated through Oil-red O staining, triacylglycerides quantification, and [(14)C] acetate incorporation into lipids. mRNA expression and protein content of SIRT1 and PPARγ were measured by RT-PCR and immunoblotting, respectively. Resveratrol-mediated SIRT1 stimuli did not induce lipogenesis and reduced the retinol-mediated fat-storing capacity in GRX. In order to support our results, we established a cell culture model of transgenic super expression of PPARγ in GRX cells (GRXPγ). Resveratrol reduced lipid droplets accumulation in GRXPγ cells. These results suggest that the PPARγ/SIRT1 ratio plays an important role in the fate of HSC. Thus, whenever the PPARγ activity is greater than SIRT1 activity the lipogenesis is enabled.
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Affiliation(s)
- Izabel Cristina Custódio de Souza
- Departamento de Morfologia, IB, Universidade Federal de Pelotas (UFPel), av. Duque de Caxias, 250, CEP 96 030 000, Pelotas, RS, Brazil.,Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul (UFRGS), rua Ramiro Barcelos, 2600-Anexo I, CEP 90035-003, Porto Alegre, RS, Brazil
| | - Leo Anderson Meira Martins
- Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul (UFRGS), rua Ramiro Barcelos, 2600-Anexo I, CEP 90035-003, Porto Alegre, RS, Brazil
| | - Mariana de Vasconcelos
- Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul (UFRGS), rua Ramiro Barcelos, 2600-Anexo I, CEP 90035-003, Porto Alegre, RS, Brazil
| | - Cleverson Moraes de Oliveira
- Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul (UFRGS), rua Ramiro Barcelos, 2600-Anexo I, CEP 90035-003, Porto Alegre, RS, Brazil
| | - Florencia Barbé-Tuana
- Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul (UFRGS), rua Ramiro Barcelos, 2600-Anexo I, CEP 90035-003, Porto Alegre, RS, Brazil
| | | | - Letícia Ferreira Pettenuzzo
- Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul (UFRGS), rua Ramiro Barcelos, 2600-Anexo I, CEP 90035-003, Porto Alegre, RS, Brazil
| | - Radovan Borojevic
- Departamento de Histologia e Embriologia, ICB, PABCAM, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, Brazil
| | - Rogério Margis
- Departamento de Biofísica, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil
| | - Regina Guaragna
- Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul (UFRGS), rua Ramiro Barcelos, 2600-Anexo I, CEP 90035-003, Porto Alegre, RS, Brazil
| | - Fátima Costa Rodrigues Guma
- Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul (UFRGS), rua Ramiro Barcelos, 2600-Anexo I, CEP 90035-003, Porto Alegre, RS, Brazil
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10
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Chang YJ, Hsu SL, Liu YT, Lin YH, Lin MH, Huang SJ, Ho JAA, Wu LC. Gallic acid induces necroptosis via TNF-α signaling pathway in activated hepatic stellate cells. PLoS One 2015; 10:e0120713. [PMID: 25816210 PMCID: PMC4376672 DOI: 10.1371/journal.pone.0120713] [Citation(s) in RCA: 36] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/11/2014] [Accepted: 01/26/2015] [Indexed: 12/16/2022] Open
Abstract
Gallic acid (3, 4, 5-trihydroxybenzoic acid, GA), a natural phenolic acid widely found in gallnuts, tea leaves and various fruits, possesses several bioactivities against inflammation, oxidation, and carcinogenicity. The beneficial effect of GA on the reduction of animal hepatofibrosis has been indicated due to its antioxidative property. However, the cytotoxicity of GA autoxidation causing cell death has also been reported. Herein, we postulated that GA might target activated hepatic stellate cells (aHSCs), the cell type responsible for hepatofibrosis, to mitigate the process of fibrosis. The molecular cytotoxic mechanisms that GA exerted on aHSCs were then analyzed. The results indicated that GA elicited aHSC programmed cell death through TNF–α–mediated necroptosis. GA induced significant oxidative stress through the suppression of catalase activity and the depletion of glutathione (GSH). Elevated oxidative stress triggered the production of TNF–α facilitating the undergoing of necroptosis through the up-regulation of key necroptotic regulatory proteins TRADD and receptor-interacting protein 3 (RIP3), and the inactivation of caspase–8. Calmodulin and calpain–1 activation were engaged, which promoted subsequent lysosomal membrane permeabilization (LMP). The TNF–α antagonist (SPD–304) and the RIP1 inhibitor (necrostatin–1, Nec–1) confirmed GA-induced TNFR1–mediated necroptosis. The inhibition of RIP1 by Nec–1 diverted the cell death from necroptosis to apoptosis, as the activation of caspase 3 and the increase of cytochrome c. Collectively, this is the first report indicating that GA induces TNF signaling–triggered necroptosis in aHSCs, which may offer an alternative strategy for the amelioration of liver fibrosis.
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Affiliation(s)
- Ya Ju Chang
- Department of Biochemical Science and Technology, National Taiwan University, Taipei, Taiwan
| | - Shih Lan Hsu
- Department of Education and Research, Taichung Veterans General Hospital, Taichung, Taiwan
| | - Yi Ting Liu
- Department of Applied Chemistry, National Chi Nan University, Puli, Taiwan
| | - Yu Hsuan Lin
- Department of Applied Chemistry, National Chi Nan University, Puli, Taiwan
| | - Ming Hui Lin
- Department of Applied Chemistry, National Chi Nan University, Puli, Taiwan
| | - Shu Jung Huang
- Department of Applied Chemistry, National Chi Nan University, Puli, Taiwan
| | - Ja-an Annie Ho
- Department of Biochemical Science and Technology, National Taiwan University, Taipei, Taiwan
| | - Li-Chen Wu
- Department of Applied Chemistry, National Chi Nan University, Puli, Taiwan
- * E-mail:
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11
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Houldsworth A, Metzner M, Shaw S, Kaminski E, Demaine AG, Cramp ME. Polymorphic differences in SOD-2 may influence HCV viral clearance. J Med Virol 2014; 86:941-7. [PMID: 24610415 DOI: 10.1002/jmv.23923] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 02/11/2014] [Indexed: 02/06/2023]
Abstract
Hepatitis C virus (HCV) is a pathogen causing chronic hepatitis, cirrhosis, and liver cancer occurring in about 3% of the world's population. Most individuals infected with HCV develop persistent viremia. Oxidative stress may play an important role in the pathogenesis of a number of diseases including HCV infection and diabetes mellitus. Polymorphisms in the antioxidant genes may determine cellular oxidative stress levels as a primary pathogenic role in HCV and/or in its complications. Patients with HCV and normal, healthy controls were investigated for a superoxide dismutase (SOD-2) polymorphism in the mitochondrial targeting sequence with Ala/Val (C-9T) substitution. Polymorphisms in antioxidant gene SOD-2 were carried out by PCR, restriction fragment length polymorphism assays and by polyacrylamide gel electrophoresis. For the SOD-2 polymorphism, the RNA positive group showed a higher percentage of "CT" genotype than the RNA negative group (89.3% vs. 66.1%, P = 0.001, χ(2) = 11.9). The RNA negative group had more TT genotypes than the RNA positive group (27.4% vs. 6.80%, P = 0.01, χ(2) = 11.6). The exposed uninfected group had an increased frequency of the "CT" genotype (86.2% vs. 66.1%, P = 0.02, χ(2) = 5.5). The RNA positives had a higher frequency of the "CT" from the normal controls (72.1% vs. 89.2%, P = 0.005, χ(2) = 7.8).
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Affiliation(s)
- Annwyne Houldsworth
- Hepatology and Molecular Medicine Research Groups, Plymouth University Peninsula School of Medicine and Dentistry (PU-PSMD), Plymouth, United Kingdom
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12
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de Mesquita FC, Bitencourt S, Caberlon E, da Silva GV, Basso BS, Schmid J, Ferreira GA, de Oliveira FDS, de Oliveira JR. Fructose-1,6-bisphosphate induces phenotypic reversion of activated hepatic stellate cell. Eur J Pharmacol 2013; 720:320-5. [DOI: 10.1016/j.ejphar.2013.09.067] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/16/2013] [Revised: 09/13/2013] [Accepted: 09/29/2013] [Indexed: 12/27/2022]
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13
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Cytotoxic effect of amphotericin B in a myofibroblast cell line. Toxicol In Vitro 2013; 27:2105-9. [PMID: 23988732 DOI: 10.1016/j.tiv.2013.08.005] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/30/2013] [Revised: 08/12/2013] [Accepted: 08/13/2013] [Indexed: 12/20/2022]
Abstract
In this study we investigate whether Amphotericin B (AmB), a widely used antifungal agent, could decrease the proliferation of a myofibroblast cell line - GRX, a model of activated hepatic stellate cells (HSC). Three different hepatic cell lines (GRX, Hep G2 and ARL-6) were treated with two concentrations of AmB (1.25 μg/mL or 2.50 μg/mL). Cytotoxicity was assessed by MTT assay. The effects of AmB on GRX migration was evaluated by Wound-healing Assay. Cell cycle arrest was investigated by flow cytometry. Apoptosis and autophagy were analyzed by Caspase 3 and LC3 immunostaining, respectively. Treatment with AmB 1.25 or 2.50 μg/mL showed a decrease in viability of GRX cells. This decrease was not observed for Hep G2 or ARL-6 in any of the two AmB concentrations tested. GRX cells treated with 1.25 μg/mL AmB were unable to close the wound after 96 h. Cell cycle analysis showed an increase in sub-G1 population and a decrease in G2/M population in AmB-treated cells. In addition, AmB-treated GRX cells showed increased expression of LC-3 and Caspase-3 by immunohistochemistry, suggesting an increase in both autophagy and apoptosis. Here we show that AmB is cytotoxic for GRX cells, a model of activated HSC, but not for hepatic lineages HepG2 and ARL6.
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14
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Dunning S, Ur Rehman A, Tiebosch MH, Hannivoort RA, Haijer FW, Woudenberg J, van den Heuvel FAJ, Buist-Homan M, Faber KN, Moshage H. Glutathione and antioxidant enzymes serve complementary roles in protecting activated hepatic stellate cells against hydrogen peroxide-induced cell death. Biochim Biophys Acta Mol Basis Dis 2013; 1832:2027-34. [PMID: 23871839 DOI: 10.1016/j.bbadis.2013.07.008] [Citation(s) in RCA: 88] [Impact Index Per Article: 7.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/29/2013] [Revised: 07/05/2013] [Accepted: 07/09/2013] [Indexed: 12/13/2022]
Abstract
BACKGROUND In chronic liver disease, hepatic stellate cells (HSCs) are activated, highly proliferative and produce excessive amounts of extracellular matrix, leading to liver fibrosis. Elevated levels of toxic reactive oxygen species (ROS) produced during chronic liver injury have been implicated in this activation process. Therefore, activated hepatic stellate cells need to harbor highly effective anti-oxidants to protect against the toxic effects of ROS. AIM To investigate the protective mechanisms of activated HSCs against ROS-induced toxicity. METHODS Culture-activated rat HSCs were exposed to hydrogen peroxide. Necrosis and apoptosis were determined by Sytox Green or acridine orange staining, respectively. The hydrogen peroxide detoxifying enzymes catalase and glutathione-peroxidase (GPx) were inhibited using 3-amino-1,2,4-triazole and mercaptosuccinic acid, respectively. The anti-oxidant glutathione was depleted by L-buthionine-sulfoximine and repleted with the GSH-analogue GSH-monoethylester (GSH-MEE). RESULTS Upon activation, HSCs increase their cellular glutathione content and GPx expression, while MnSOD (both at mRNA and protein level) and catalase (at the protein level, but not at the mRNA level) decreased. Hydrogen peroxide did not induce cell death in activated HSCs. Glutathione depletion increased the sensitivity of HSCs to hydrogen peroxide, resulting in 35% and 75% necrotic cells at 0.2 and 1mmol/L hydrogen peroxide, respectively. The sensitizing effect was abolished by GSH-MEE. Inhibition of catalase or GPx significantly increased hydrogen peroxide-induced apoptosis, which was not reversed by GSH-MEE. CONCLUSION Activated HSCs have increased ROS-detoxifying capacity compared to quiescent HSCs. Glutathione levels increase during HSC activation and protect against ROS-induced necrosis, whereas hydrogen peroxide-detoxifying enzymes protect against apoptotic cell death.
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Affiliation(s)
- Sandra Dunning
- Department of Gastroenterology and Hepatology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands
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15
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Resveratrol Induces Pro-oxidant Effects and Time-Dependent Resistance to Cytotoxicity in Activated Hepatic Stellate Cells. Cell Biochem Biophys 2013; 68:247-57. [DOI: 10.1007/s12013-013-9703-8] [Citation(s) in RCA: 43] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/26/2022]
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16
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Copper ions stimulate the proliferation of hepatic stellate cells via oxygen stress in vitro. ACTA ACUST UNITED AC 2013; 33:75-80. [DOI: 10.1007/s11596-013-1074-2] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2012] [Indexed: 12/19/2022]
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17
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Li JF, Zheng SJ, Duan ZP. Liver fibrosis: Role of oxidative stress and therapeutic countermeasures. Shijie Huaren Xiaohua Zazhi 2013; 21:1573. [DOI: 10.11569/wcjd.v21.i17.1573] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
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18
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Bragança de Moraes CM, Melo DADS, Santos RCV, Bitencourt S, Mesquita FC, Santos de Oliveira FD, Rodrıguez-Carballo E, Bartrons R, Rosa JL, Ventura FP, Rodrigues de Oliveira J. Antiproliferative effect of catechin in GRX cells. Biochem Cell Biol 2012; 90:575-84. [DOI: 10.1139/o2012-010] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022] Open
Abstract
The phenolic compounds present in cocoa seeds have been studied regarding health benefits, such as antioxidant and anti-inflammatory activities. Fibrosis is a wound healing response that occurs in almost all patients with chronic liver injury. A large number of cytokines and soluble intercellular mediators are related to changes in the behavior and phenotype of the hepatic stellate cell (HSC) that develop a fibrogenic and contractile phenotype leading to the development of fibrosis. The objective of this study was to assess the catechin effect in GRX liver cells in activities such as cell growth and inflammation. The GRX cells treatment with catechin induced a significant decrease in cell growth. This mechanism does not occur by apoptosis or even by autophagy because there were no alterations in expression of caspase 3 and PARP (apoptosis), and LC3 (autophagy). The expression of p27 and p53 proteins, regulators of the cell cycle, showed increased expression, while COX-2 and IL-6 mRNA showed a significant decrease in expression. This study shows that catechin decreases cell growth in GRX cells and, probably, this decrease does not occur by apoptosis or autophagy but through an anti-inflammatory effect and cell cycle arrest. Catechin also significantly decreased the production of TGF-β by GRX cells, showing a significant antifibrotic effect.
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Affiliation(s)
- Cristina Machado Bragança de Moraes
- Laboratório de Biofísica Celular e Inflamação, Pontifícia Universidade Católica do Rio Grande do Sul, PUCRS, Avenida Ipiranga 6681, prédio 12, CEP: 90619-900 Porto Alegre, Rio Grande do Sul, Brazil
| | - Denizar Alberto da Silva Melo
- Laboratório de Biofísica Celular e Inflamação, Pontifícia Universidade Católica do Rio Grande do Sul, PUCRS, Avenida Ipiranga 6681, prédio 12, CEP: 90619-900 Porto Alegre, Rio Grande do Sul, Brazil
| | - Roberto Christ Vianna Santos
- Laboratório de Biofísica Celular e Inflamação, Pontifícia Universidade Católica do Rio Grande do Sul, PUCRS, Avenida Ipiranga 6681, prédio 12, CEP: 90619-900 Porto Alegre, Rio Grande do Sul, Brazil
| | - Shanna Bitencourt
- Laboratório de Biofísica Celular e Inflamação, Pontifícia Universidade Católica do Rio Grande do Sul, PUCRS, Avenida Ipiranga 6681, prédio 12, CEP: 90619-900 Porto Alegre, Rio Grande do Sul, Brazil
| | - Fernanda Cristina Mesquita
- Laboratório de Biofísica Celular e Inflamação, Pontifícia Universidade Católica do Rio Grande do Sul, PUCRS, Avenida Ipiranga 6681, prédio 12, CEP: 90619-900 Porto Alegre, Rio Grande do Sul, Brazil
| | | | - Edgardo Rodrıguez-Carballo
- Departament de Ciències Fisiològiques II, IDIBELL, Universitat de Barcelona, Campus de Bellvitge, Barcelona, Spain
| | - Ramon Bartrons
- Departament de Ciències Fisiològiques II, IDIBELL, Universitat de Barcelona, Campus de Bellvitge, Barcelona, Spain
| | - Jose Luis Rosa
- Departament de Ciències Fisiològiques II, IDIBELL, Universitat de Barcelona, Campus de Bellvitge, Barcelona, Spain
| | - Francesc Pujol Ventura
- Departament de Ciències Fisiològiques II, IDIBELL, Universitat de Barcelona, Campus de Bellvitge, Barcelona, Spain
| | - Jarbas Rodrigues de Oliveira
- Laboratório de Biofísica Celular e Inflamação, Pontifícia Universidade Católica do Rio Grande do Sul, PUCRS, Avenida Ipiranga 6681, prédio 12, CEP: 90619-900 Porto Alegre, Rio Grande do Sul, Brazil
- Hospital de Clínicas de Porto Alegre, Research Center, Ramiro Barcelos 2.350 CEP 90035-90 Porto Alegre / RS
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Bian Z, Ma X. Liver fibrogenesis in non-alcoholic steatohepatitis. Front Physiol 2012; 3:248. [PMID: 22934006 PMCID: PMC3429026 DOI: 10.3389/fphys.2012.00248] [Citation(s) in RCA: 31] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/14/2012] [Accepted: 06/17/2012] [Indexed: 12/13/2022] Open
Abstract
Non-alcoholic fatty liver disease (NAFLD) is emerging as one of the most common chronic liver diseases in developed western countries. Non-alcoholic steatohepatitis (NASH) is the most severe form of NAFLD, and can progress to more severe forms of liver disease, including fibrosis, cirrhosis, and even hepatocellular carcinoma. The activation of hepatic stellate cells plays a critical role in NASH-related fibrogenesis. Multiple factors, such as insulin resistance, oxidative stress, pro-inflammatory cytokines and adipokines, and innate immune responses, are known to contribute to the development of NASH-related fibrogenesis. Furthermore, these factors may share synergistic interactions, which could contribute to the process of liver fibrosis. Given the complex etiology of NASH, combined treatment regimes that target these different factors provide potential treatment strategies for NASH-related liver fibrosis.
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Affiliation(s)
- Zhaolian Bian
- Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Renji Hospital, Shanghai Jiao-Tong University School of Medicine Shanghai, China
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20
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Ali G, Mohsin S, Khan M, Nasir GA, Shams S, Khan SN, Riazuddin S. Nitric oxide augments mesenchymal stem cell ability to repair liver fibrosis. J Transl Med 2012; 10:75. [PMID: 22533821 PMCID: PMC3419634 DOI: 10.1186/1479-5876-10-75] [Citation(s) in RCA: 55] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/22/2012] [Accepted: 04/25/2012] [Indexed: 12/15/2022] Open
Abstract
Background Liver fibrosis is a major health problem worldwide and poses a serious obstacle for cell based therapies. Mesenchymal stem cells (MSCs) are multipotent and important candidate cells for future clinical applications however success of MSC therapy depends upon their homing and survival in recipient organs. This study was designed to improve the repair potential of MSCs by transplanting them in sodium nitroprusside (SNP) pretreated mice with CCl4 induced liver fibrosis. Methods SNP 100 mM, a nitric oxide (NO) donor, was administered twice a week for 4 weeks to CCl4-injured mice. MSCs were isolated from C57BL/6 wild type mice and transplanted in the left lateral lobe of the liver in experimental animals. After 4 weeks, animals were sacrificed and liver improvement was analyzed. Analysis of fibrosis by qRT-PCR and sirius red staining, homing, bilirubin and alkaline phosphatase (ALP) serum levels between different treatment groups were compared to control. Results Liver histology demonstrated enhanced MSCs homing in SNP-MSCs group compared to MSCs group. The gene expression of fibrotic markers; αSMA, collagen 1α1, TIMP, NFκB and iNOS was down regulated while cytokeratin 18, albumin and eNOS was up-regulated in SNP-MSCs group. Combine treatment sequentially reduced fibrosis in SNP-MSCs treated liver compared to the other treatment groups. These results were also comparable with reduced serum levels of bilirubin and ALP observed in SNP-MSCs treated group. Conclusion This study demonstrated that NO effectively augments MSC ability to repair liver fibrosis induced by CCl4 in mice and therefore is a better treatment regimen to reduce liver fibrosis.
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Affiliation(s)
- Gibran Ali
- National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan
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Singal AK, Jampana SC, Weinman SA. Antioxidants as therapeutic agents for liver disease. Liver Int 2011; 31:1432-48. [PMID: 22093324 PMCID: PMC3228367 DOI: 10.1111/j.1478-3231.2011.02604.x] [Citation(s) in RCA: 156] [Impact Index Per Article: 11.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/13/2010] [Accepted: 06/15/2011] [Indexed: 12/12/2022]
Abstract
Oxidative stress is commonly associated with a number of liver diseases and is thought to play a role in the pathogenesis of chronic hepatitis C, alcoholic liver disease, non-alcoholic steatohepatitis (NASH), haemochromatosis and Wilson's disease. Antioxidant therapy has thus been considered to have the possibility of beneficial effects in the management of these liver diseases. Despite this promise, antioxidants have produced mixed results in a number of clinical trials of efficacy. This review summarizes the results of clinical trials of antioxidants as sole or adjuvant therapy of chronic hepatitis C, alcoholic liver disease and non-alcoholic steatohepatitis (NASH). Overall, the most promising results to date are for vitamin E therapy of NASH but some encouraging results have been obtained with antioxidant therapy of acute alcoholic hepatitis as well. Despite evidence for small reductions of serum alanine aminotransferase, there is as yet no convincing evidence that antioxidant therapy itself is beneficial to patients with chronic hepatitis C. Problems such as small sample size, short follow up duration, inadequate endpoints, failure to demonstrate tissue delivery and antioxidant efficacy, and heterogeneous nature of the 'antioxidant' compounds used have complicated interpretation of results of the clinical studies. These limitations and their implications for future trial design are discussed.
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Affiliation(s)
- Ashwani K. Singal
- Department of Internal Medicine, University of Texas Medical Branch; Galveston, TX
| | - Sarat C. Jampana
- Department of Internal Medicine, University of Texas Medical Branch; Galveston, TX
| | - Steven A. Weinman
- Department of Internal Medicine; University of Kansas Medical Center; Kansas City, KS
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Stefano JT, Cogliati B, Santos F, Lima VMR, Mazo DC, Matte U, Alvares-da-Silva MR, Silveira TR, Carrilho FJ, Oliveira CPMS. S-Nitroso-N-acetylcysteine induces de-differentiation of activated hepatic stellate cells and promotes antifibrotic effects in vitro. Nitric Oxide 2011; 25:360-365. [PMID: 21820071 DOI: 10.1016/j.niox.2011.07.001] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/10/2011] [Revised: 06/22/2011] [Accepted: 07/04/2011] [Indexed: 01/24/2023]
Abstract
Nitric oxide (NO) has been shown to act as a potent antifibrogenic agent by decreasing myofibroblast differentiation. S-Nitroso-N-acetylcysteine (SNAC), a NO donor, attenuates liver fibrosis in rats, but the cellular and molecular mechanisms on liver myofibroblast-like phenotype still remain unknown. Here, we investigate the antifibrotic effects of SNAC on hepatic stellate cells, the major fibrogenic cell type in the liver. A murine GRX cell line was incubated with SNAC (100μM) or vehicle (control group) for 72h. Cell viability was measured by MTT colorimetric assay and the conversion of myofibroblast into quiescent fat-storing cell phenotype was evaluated by Oil-Red-O staining. TGFβ-1, TIMP-1, and MMP-13 levels were measure in the supernatant by ELISA. Profibrogenic- and fibrolytic-related gene expression was quantified using real-time qPCR. SNAC induced phenotype conversion of myofibroblast-like phenotype into quiescent cells. SNAC decreased gene and protein expression of TGFβ-1 and MMP-2 compared to control groups. Besides, SNAC down-regulated profibrogenic molecules and up-regulated MMP-13 gene expression, which plays a key role in the degradation of interstitial collagen in liver fibrosis. In conclusion, these findings demonstrate that SNAC efficiently can modulate the activation and functionality of murine hepatic stellate cells and could be considered as an antifibrotic treatment to human liver fibrosis.
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Affiliation(s)
- J T Stefano
- Department of Gastroenterology, University of São Paulo School of Medicine, São Paulo, Brazil
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23
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Madill J, Arendt B, Aghdassi E, Chow C, Guindi M, Therapondos G, Lilly L, Allard J. Oxidative Stress and Nutritional Factors in Hepatitis C Virus–Positive Liver Recipients, Controls, and Hepatitis C Virus–Positive Nontransplant Patients. Transplant Proc 2010; 42:1744-9. [DOI: 10.1016/j.transproceed.2010.03.141] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/05/2009] [Accepted: 03/10/2010] [Indexed: 01/25/2023]
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24
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Andrade CMB, Wink MR, Margis R, Borojevic R, Battastini AMO, Guma FCR. Changes in E-NTPDase 3 expression and extracellular nucleotide hydrolysis during the myofibroblast/lipocyte differentiation. Mol Cell Biochem 2010; 339:79-87. [PMID: 20058055 DOI: 10.1007/s11010-009-0371-3] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/09/2009] [Accepted: 12/16/2009] [Indexed: 01/07/2023]
Abstract
Hepatic stellate cells (HSC) play a critical role in the development and maintenance of liver fibrosis. HSC are lipocytes that displayed the capacity to develop into myofibroblast-like cells. Ecto-nucleoside triphosphate diphosphohydrolases (E-NTPDases) regulate the concentration of extracellular nucleotides, signaling molecules that play a role in the pathogenesis of hepatic fibrosis. In the present study, we identified and compared the expressions of E-NTPDase family members in two different phenotypes of the mouse hepatic stellate cell line (GRX) and evaluated the nucleotide hydrolysis by these cells. We show that both phenotypes of GRX cell line expressed NTPDase 3 and 5. However, only activated cells expressed NTPDase 6. In quiescent-like cells, the hydrolysis of triphosphonucleosides was significantly higher, and was related to an increase in Entpd3 mRNA expression. The diphosphonucleosides were hydrolyzed at a similar rate by two phenotypes of GRX cells. We suggest that up-regulation of Entpd3 mRNA expression modulates the extracellular concentration of nucleotides/nucleosides and affect P2-receptor signaling differently in quiescent-like cells and may play a role in the regulation of HSC functions.
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Affiliation(s)
- Cláudia M B Andrade
- Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, UFRGS, Rua Ramiro Barcelos, 2600-anexo, Porto Alegre, RS CEP 90035-003, Brazil
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25
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Dunning S, Hannivoort RA, de Boer JF, Buist-Homan M, Faber KN, Moshage H. Superoxide anions and hydrogen peroxide inhibit proliferation of activated rat stellate cells and induce different modes of cell death. Liver Int 2009; 29:922-32. [PMID: 19386027 DOI: 10.1111/j.1478-3231.2009.02004.x] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/13/2023]
Abstract
BACKGROUND In chronic liver injury, hepatic stellate cells (HSCs) proliferate and produce excessive amounts of connective tissue causing liver fibrosis and cirrhosis. Oxidative stress has been implicated as a driving force of HSC activation and proliferation, although contradictory results have been described. AIM To determine the effects of oxidative stress on activated HSC proliferation, survival and signalling pathways. METHODS Serum-starved culture-activated rat HSCs were exposed to the superoxide anion donor menadione (5-25 micromol/L) or hydrogen peroxide (0.2-5 mmol/L). Haem oxygenase-1 mRNA expression, glutathione status, cell death, phosphorylation of mitogen-activated protein (MAP) kinases and proliferation were investigated. RESULTS Menadione induced apoptosis in a dose- and time-dependent, but caspase-independent manner. Hydrogen peroxide induced necrosis only at extremely high concentrations. Both menadione and hydrogen peroxide activated Jun N-terminal kinase (JNK) and p38. Hydrogen peroxide also activated extracellular signal-regulated protein. Menadione, but not hydrogen peroxide, reduced cellular glutathione levels. Inhibition of JNK or supplementation of glutathione reduced menadione-induced apoptosis. Non-toxic concentrations of menadione or hydrogen peroxide inhibited platelet-derived growth factor- or/and serum-induced proliferation. CONCLUSION Reactive oxygen species (ROS) inhibit HSC proliferation and promote HSC cell death in vitro. Different ROS induce different modes of cell death. Superoxide anion-induced HSC apoptosis is dependent on JNK activation and glutathione status.
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Affiliation(s)
- Sandra Dunning
- Department of Gastroenterology and Hepatology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
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26
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Trappoliere M, Caligiuri A, Schmid M, Bertolani C, Failli P, Vizzutti F, Novo E, di Manzano C, Marra F, Loguercio C, Pinzani M. Silybin, a component of sylimarin, exerts anti-inflammatory and anti-fibrogenic effects on human hepatic stellate cells. J Hepatol 2009; 50:1102-11. [PMID: 19398228 DOI: 10.1016/j.jhep.2009.02.023] [Citation(s) in RCA: 174] [Impact Index Per Article: 10.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/25/2008] [Revised: 12/19/2008] [Accepted: 02/04/2009] [Indexed: 02/06/2023]
Abstract
BACKGROUND/AIMS Hepatic fibrogenesis, a consequence of chronic liver tissue damage, is characterized by activation of the hepatic stellate cells (HSC). Silybin has been shown to exert anti-fibrogenic effects in animal models. However, scant information is available on the fine cellular and molecular events responsible for this effect. The aim of this study was to assess the mechanisms regulating the anti-fibrogenic and anti-inflammatory activity of Silybin. METHODS Experiments were performed on HSC isolated from human liver and activated by culture on plastic. RESULTS Silybin was able to inhibit dose-dependently (25-50 microM) growth factor-induced pro-fibrogenic actions of activated human HSC, including cell proliferation (P < 0.001), cell motility (P < 0.001), and de novo synthesis of extracellular matrix components (P < 0.05). Silybin (25-50 microM), inhibited the IL-1-induced synthesis of MCP-1 (P < 0.01) and IL-8 (P < 0.01) showing a potent anti-inflammatory activity. Silybin exerts its effects by directly inhibiting the ERK, MEK and Raf phosphorylation, reducing the activation of NHE1 (Na+/H+ exchanger, P < 0.05) and the IkBalpha phosphorylation. In addition, Silybin was confirmed to act as a potent anti-oxidant agent. CONCLUSION The results of the study provide molecular insights into the potential therapeutic action of Silybin in chronic liver disease. This action seems to be mostly related to a marked inhibition of the production of pro-inflammatory cytokines, a clear anti-oxidant effect and a reduction of the direct and indirect pro-fibrogenic potential of HSC.
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Affiliation(s)
- Marco Trappoliere
- Dipartimento di Medicina Interna, Università degli Studi di Firenze, Viale G.B. Morgagni, 85, 50134 Florence, Italy
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27
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Andrade CMB, Wink MR, Margis R, Borojevic R, Battastini AMO, Guma FCR. Activity and expression of ecto-nucleotide pyrophosphate/phosphodiesterases in a hepatic stellate cell line. Mol Cell Biochem 2009; 325:179-85. [PMID: 19194664 DOI: 10.1007/s11010-009-0032-6] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/12/2008] [Accepted: 01/15/2009] [Indexed: 01/04/2023]
Abstract
Nucleotides and nucleosides represent an important and ubiquitous class of molecules that interact with specific receptors, regulate a variety of activities within the liver, and play a role in the pathogenesis of hepatic fibrosis. Ecto-nucleotide pyrophosphatase/phosphodiesterases (E-NPPs) are ecto-enzymes that are located on the cell surface. NPP1, NPP2, and NPP3 (abbreviated as NPP1-3 hereafter) have been implicated in the hydrolysis of nucleotides; together with other ecto-nucleotidases, they control the events induced by extracellular nucleotides. We have identified and compared the expression of E-NPP family members in two different phenotypes of the mouse hepatic stellate cell line (GRX). In quiescent-like hepatic stellate cells (HSCs), E-NPP activity was significantly higher, NPP2 mRNA expression decreased and NPP3 mRNA increased. The differential NPP activity and expression in two phenotypes of GRX cells suggests that they are involved in the regulation of extracellular nucleotide metabolism in HSCs. However, the role of E-NPPs in the liver remains to be clarified.
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Affiliation(s)
- Cláudia M B Andrade
- Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, UFRGS, Rua Ramiro Barcelos, 2600-anexo, Porto Alegre, CEP 90035-003, RS, Brazil
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28
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Abstract
Uncontrolled production of collagen I is the main feature of liver fibrosis. Following a fibrogenic stimulus such as alcohol, hepatic stellate cells (HSC) transform into an activated collagen-producing cell. In alcoholic liver disease, numerous changes in gene expression are associated with HSC activation, including the induction of several intracellular signaling cascades, which help maintain the activated phenotype and control the fibrogenic and proliferative state of the cell. Detailed analyses for understanding the molecular basis of the collagen I gene regulation have revealed a complex process involving reactive oxygen species (ROS) as key mediators. Less is known, however, about the contribution of reactive nitrogen species (RNS). In addition, a series of cytokines, growth factors, and chemokines, which activate extracellular matrix (ECM)-producing cells through paracrine and autocrine loops, contribute to the fibrogenic response.
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Affiliation(s)
- R. Urtasun
- Mount Sinai School of Medicine, Box 1123, Department of Medicine/Division of Liver Diseases, 1425 Madison Avenue, Room 11-76, New York, NY 10029, USA
| | - L. Conde de la Rosa
- Mount Sinai School of Medicine, Box 1123, Department of Medicine/Division of Liver Diseases, 1425 Madison Avenue, Room 11-76, New York, NY 10029, USA
| | - N. Nieto
- Mount Sinai School of Medicine, Box 1123, Department of Medicine/Division of Liver Diseases, 1425 Madison Avenue, Room 11-76, New York, NY 10029, USA
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29
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Voltan S, Martines D, Elli M, Brun P, Longo S, Porzionato A, Macchi V, D'Incà R, Scarpa M, Palù G, Sturniolo GC, Morelli L, Castagliuolo I. Lactobacillus crispatus M247-derived H2O2 acts as a signal transducing molecule activating peroxisome proliferator activated receptor-gamma in the intestinal mucosa. Gastroenterology 2008; 135:1216-27. [PMID: 18722375 DOI: 10.1053/j.gastro.2008.07.007] [Citation(s) in RCA: 76] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/04/2007] [Revised: 06/26/2008] [Accepted: 07/03/2008] [Indexed: 12/02/2022]
Abstract
BACKGROUND & AIMS Accumulating evidence indicates that the peroxisome proliferator activated receptor (PPAR)-gamma is a major player in maintaining intestinal mucosa homeostasis, but whether PPAR-gamma is directly involved in probiotic-mediated effects and the molecular events involved in its activation are not known. METHODS We investigated the role of PPAR-gamma in the immunomodulatory effects of Lactobacillus crispatus M247 on intestinal epithelial cells (IEC) and the role of probiotic-derived H(2)O(2) on PPAR-gamma activity. RESULTS L crispatus M247 supplementation in mice significantly increased PPAR-gamma levels and transcriptional activity in the colonic mucosa. L crispatus M247 induced PPAR-gamma nuclear translocation and enhanced transcriptional activity in epithelial (CMT-93) cells, as demonstrated by the increased luciferase activity of a PPAR-gamma-responsive element, PPAR-gamma-responsive gene up-regulation, and reduced activity of an nuclear factor-kappaB-responsive element. Pharmacologic PPAR-gamma inhibition or silencing by small interfering RNA cancelled the L crispatus M247-mediated effects in CMT-93 cells. Because Lactobacillus strains producing little H(2)O(2) failed to activate PPAR-gamma, we investigated the role of L crispatus M247-derived H(2)O(2) in PPAR-gamma activation. L crispatus M247 induced a transient rise in intracellular H(2)O(2) and PPAR-gamma transcriptional activity was cancelled by antioxidant or H(2)O(2) scavenger. Toll-like receptor (TLR)-2 was not required for PPAR-gamma up-regulation mediated by L crispatus M247 in mice, although the protective effects of L crispatus M247 on dextran sodium sulfate-induced colitis were less pronounced in TLR-2(-/-) mice. CONCLUSIONS L crispatus M247 uses H(2)O(2) as a signal transducing molecule to induce PPAR-gamma activation in IEC, directly modulating epithelial cell responsiveness to inflammatory stimuli.
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Affiliation(s)
- Sandra Voltan
- Department of Histology, Microbiology and Medical Biotechnologies, University of Padua, Italy
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30
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Souza IC, Martins LAM, Coelho BP, Grivicich I, Guaragna RM, Gottfried C, Borojevic R, Guma FCR. Resveratrol inhibits cell growth by inducing cell cycle arrest in activated hepatic stellate cells. Mol Cell Biochem 2008; 315:1-7. [PMID: 18454344 DOI: 10.1007/s11010-008-9781-x] [Citation(s) in RCA: 37] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/01/2008] [Accepted: 04/22/2008] [Indexed: 12/13/2022]
Abstract
Resveratrol (RSV) exerts anti-proliferative and pro-apoptotic actions in different cell lines. Hepatic stellate cells (HSCs) are major fibrogenic cell types that contribute to collagen accumulation during chronic liver disease. In the present study, the inhibitory effects of RSV on cell proliferation, cell cycle, and apoptosis were evaluated in the mouse hepatic stellate cell line GRX. Cells treated with 1 nM-1 muM of RSV demonstrated a decrease in cell growth of about 35% after 5 days. GRX cells, treated with RSV (100 nM or 1 muM), were analyzed by flow cytometry; RSV induced an increase in the number of GRX cells in the S- and sub-G1 phases. The increase in sub-G1 phase cells and the nuclear condensation and fragmentation shown by DAPI staining identified a possible pro-apoptotic effect of RSV on GRX cells. Furthermore, the RSV anti-proliferative effects could be explained by an S-phase accumulation caused by a decrease in the progression through the cell cycle or an inhibition of S or G2 phase transition. It is notable that these RSV actions are mediated at nanomolar levels, compatible with the concentrations of free RSV in biological fluids after ingestion of polyphenol-rich foods, suggesting a possible effect of these foods as an adjuvant treatment in chronic liver diseases.
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Affiliation(s)
- Izabel C Souza
- Departamento de Bioquímica, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600-anexo, CEP 90035-003, Porto Alegre, RS, Brazil
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31
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Moreira RK. Hepatic stellate cells and liver fibrosis. Arch Pathol Lab Med 2007; 131:1728-34. [PMID: 17979495 DOI: 10.5858/2007-131-1728-hscalf] [Citation(s) in RCA: 324] [Impact Index Per Article: 18.0] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 04/24/2007] [Indexed: 11/06/2022]
Abstract
Substantial evidence now exists to recognize hepatic stellate cells (HSCs) as the main matrix-producing cells in the process of liver fibrosis. Liver injury of any etiology will ultimately lead to activation of HSCs, which undergo transdifferentiation to fibrogenic myofibroblast-like cells. Quantitative analysis of HSC activation by immunohistochemistry has been shown to be useful in predicting the rate of progression of liver fibrosis in some clinical situations. In the activation process, transforming growth factor beta is thought to be the main mediator of fibrogenesis and platelet-derived growth factor is the major inducer of HSC proliferation. Different platelet-derived growth factor and transforming growth factor beta inhibitors have been shown to effectively prevent liver fibrosis in animal models and represent promising therapeutic agents for humans.
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Affiliation(s)
- Roger Klein Moreira
- Department of Pathology and Laboratory Medicine, Emory University School of Medicine, 1364 Clifton Rd NE, Atlanta, GA 30322, USA.
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32
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Guimarães ELM, Franceschi MFS, Andrade CMB, Guaragna RM, Borojevic R, Margis R, Bernard EA, Guma FCR. Hepatic stellate cell line modulates lipogenic transcription factors. Liver Int 2007; 27:1255-64. [PMID: 17919238 DOI: 10.1111/j.1478-3231.2007.01578.x] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/10/2023]
Abstract
BACKGROUND/AIMS Pre-adipocyte differentiation into adipocyte is a terminal differentiation process triggered by a cascade of transcription factors. Conversely, hepatic stellate cells (HSC) can switch between lipid storing and the myofibroblast phenotype in association with liver fibrotic processes. Here, adipogenic/lipogenic-related transcription factors and downstream-regulated genes were evaluated in a murine HSC cell line. GRX-HSC cells are transitional myofibroblasts that differentiate into lipocytes following retinol or indomethacin treatment. METHODS/RESULTS Specific mRNAs were quantified by a real-time polymerase chain reaction after 24 h or 7 days of cell culture with indomethacin or retinol. Proliferator-activated receptorgamma and Pex16 transcripts were increased either by retinol or indomethacin. Retinol induced a minor increase in C/enhancer binding proteinalpha transcripts, while only indomethacin increased adipsin transcripts. CONCLUSIONS Our results showed that the myofibroblast to lipocyte phenotype switch follows partially different transcriptional pathways, according to the effector. Retinol induces lipid synthesis and storage without affecting characteristic adipocytic genes, while indomethacin treatment restores the lipocytic phenotype with increased adipisin expression.
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33
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Andrade CMB, Roesch GC, Wink MR, Guimarães ELM, Souza LF, Jardim FR, Guaragna RM, Bernard EA, Margis R, Borojevic R, Battastini AMO, Guma FCR. Activity and expression of ecto-5'-nucleotidase/CD73 are increased during phenotype conversion of a hepatic stellate cell line. Life Sci 2007; 82:21-9. [PMID: 18037449 DOI: 10.1016/j.lfs.2007.10.003] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/21/2007] [Revised: 10/10/2007] [Accepted: 10/16/2007] [Indexed: 01/08/2023]
Abstract
Hepatic stellate cells (HSC) play a crucial role in the development of liver fibrosis and are important targets in liver disease therapy. Adenosine acts as an extracellular signaling molecule in various tissues and in liver this nucleoside exerts protective effects. Ecto-5'-nucleotidase/CD73 is a marker for the plasma membrane and is considered to be a key enzyme in the generation of adenosine in the extracellular medium, by transforming AMP into adenosine. In addition, adenosine production from AMP is also catalyzed by alkaline phosphatase. We compared the extracellular metabolism of AMP and transcriptional levels of the ecto-5'-nucleotidase/CD73 and tissue non-specific alkaline phosphatase (TNALP) in activated and quiescent HSC of the mouse hepatic stellate cell line GRX. This cell line expresses a myofibroblast phenotype in basal medium and both retinol and indomethacin treatment induced a phenotypic change of GRX cells to quiescent HSC. Ecto-5'-nucleotidase activity and its mRNA expression were found to be higher in quiescent HSC than in activated HSC. During phenotype conversion, mediated by retinol, the AMP decay was accelerated with adenosine accumulation in extracellular medium, likely due to the decrease in adenosine deaminase activity also observed in quiescent HSC. The treatment with retinol also involves transcriptional activation of TNALP. Taken together, these data suggest that ecto-5'-nucleotidase-dependent adenosine generation may play a role in the regulation of quiescent HSC functions.
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Affiliation(s)
- Cláudia M B Andrade
- Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, UFRGS. Porto Alegre, RS, Brazil
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34
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de Souza LF, Barreto F, da Silva EG, Andrades ME, Guimarães ELM, Behr GA, Moreira JCF, Bernard EA. Regulation of LPS stimulated ROS production in peritoneal macrophages from alloxan-induced diabetic rats: involvement of high glucose and PPARgamma. Life Sci 2007; 81:153-9. [PMID: 17532345 DOI: 10.1016/j.lfs.2007.04.035] [Citation(s) in RCA: 37] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/10/2007] [Revised: 04/15/2007] [Accepted: 04/27/2007] [Indexed: 01/21/2023]
Abstract
An increased occurrence of long term bacterial infections is common in diabetic patients. Bacterial cell wall components are described as the main antigenic agents from these microorganisms and high blood glucose levels are suggested to be involved in altered immune response. Hyperglycemia is reported to alter macrophages response to lipopolysaccharide (LPS) and peroxisome proliferators activated receptor gamma (PPARgamma) expression. Additionally, glucose is the main metabolic fuel for reduced nicotinamide adenine dinucleotide phosphate (NADPH) production by pentose phosphate shunt. In this work, lipopolysaccharide (LPS) stimulated reactive oxygen species (ROS) and nitrite production were evaluated in peritoneal macrophages from alloxan-induced diabetic rats. Cytosolic dehydrogenases and PPARgamma expression were also investigated. LPS was ineffective to stimulate ROS and nitrite production in peritoneal macrophages from diabetic rats, which presented increased glucose-6-phosphate dehydrogenase and malate dehydrogenase activity. In RAW 264.7 macrophages, acute high glucose treatment abolished LPS stimulated ROS production, with no effect on nitrite and dehydrogenase activities. Peritoneal macrophages from alloxan-treated rats presented reduced PPARgamma expression. Treating RAW 264.7 macrophages with a PPARgamma antagonist resulted in defective ROS production in response to LPS, however, stimulated nitrite production was unaltered. In conclusion, in the present study we have reported reduced nitric oxide and reactive oxygen species production in LPS-treated peritoneal macrophages from alloxan-induced diabetic rats. The reduced production of reactive oxygen species seems to be dependent on elevated glucose levels and reduced PPARgamma expression.
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Hsiao TJ, Liao HWC, Hsieh PS, Wong RH. Risk of betel quid chewing on the development of liver cirrhosis: a community-based case-control study. Ann Epidemiol 2007; 17:479-85. [PMID: 17448681 DOI: 10.1016/j.annepidem.2006.12.007] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/23/2006] [Revised: 11/23/2006] [Accepted: 12/05/2006] [Indexed: 11/25/2022]
Abstract
PURPOSE The role of betel quid on the development of liver cirrhosis is unclear; we thus designed a community-based case-control study to evaluate the association between betel quid chewing and liver cirrhosis. METHODS A total of 42 cases of liver cirrhosis and 165 matched controls were included for analysis. Questionnaires were administered to obtain histories of betel quid chewing, alcohol consumption, smoking, and family history of liver disease. Hepatitis B surface antigen and anti-hepatitis C antibody were also determined by immunoassay. RESULTS Individuals with more betel quid chewing (more than 55 quid-years vs. less than 55 quid-years and never-chewers, matched odds ratio [OR(m)] = 2.2; 95% confidence interval [CI]: 1.0-5.0) had higher risks for liver cirrhosis. The combined effects on liver cirrhosis by betel quid chewing and the number of other risk factors, including hepatitis B virus (HBV) infection, smoking, and alcohol drinking, were also observed. When individuals with less betel quid chewing (less than 55 quid-years and never-chewers) and with no other risk factors used as a reference, betel quid chewers expressing greater betel quid chewing (more than 55 quid-years) and more risk factors of HBV infection, cigarette smoking, and habitual alcohol drinking expressed a greater risk of liver cirrhosis (OR(m) = 70.8; 95% CI: 4.0-1260.1). CONCLUSIONS Our results suggest that betel quid chewing may play an important role in the development of hepatic cirrhosis. Larger study and cohort studies would be necessary to provide further evidence regarding this finding.
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Affiliation(s)
- Tun-Jen Hsiao
- Department of Public Health, College of Health Care and Management, Chung Shan Medical University, Taichung, Taiwan
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