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Epigallocatechin-3-gallate-mediated cardioprotection by Akt/GSK-3β/caveolin signalling in H9c2 rat cardiomyoblasts
BACKGROUND: Epigallocatechin-3-gallate (EGCg) with its potent anti-oxidative capabilities is known for its beneficial effects ameliorating oxidative injury to cardiac cells. Although studies have provided convincing evidence to support the cardioprotective effects of EGCg, it remains unclear whether...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3871020/ https://www.ncbi.nlm.nih.gov/pubmed/24251870 http://dx.doi.org/10.1186/1423-0127-20-86 |
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author | Hsieh, Shih-Ron Hsu, Chen-Sen Lu, Chen-Hua Chen, Wei-Cheng Chiu, Chun-Hwei Liou, Ying-Ming |
author_facet | Hsieh, Shih-Ron Hsu, Chen-Sen Lu, Chen-Hua Chen, Wei-Cheng Chiu, Chun-Hwei Liou, Ying-Ming |
author_sort | Hsieh, Shih-Ron |
collection | PubMed |
description | BACKGROUND: Epigallocatechin-3-gallate (EGCg) with its potent anti-oxidative capabilities is known for its beneficial effects ameliorating oxidative injury to cardiac cells. Although studies have provided convincing evidence to support the cardioprotective effects of EGCg, it remains unclear whether EGCg affect trans-membrane signalling in cardiac cells. Here, we have demonstrated the potential mechanism for cardioprotection of EGCg against H(2)O(2)-induced oxidative stress in H9c2 cardiomyoblasts. RESULTS: Exposing H9c2 cells to H(2)O(2) suppressed cell viability and altered the expression of adherens and gap junction proteins with increased levels of intracellular reactive oxygen species and cytosolic Ca(2+). These detrimental effects were attenuated by pre-treating cells with EGCg for 30 min. EGCg also attenuated H(2)O(2)-mediated cell cycle arrest at the G1-S phase through the glycogen synthase kinase-3β (GSK-3β)/β-catenin/cyclin D1 signalling pathway. To determine how EGCg targets H9c2 cells, enhanced green fluorescence protein (EGFP) was ectopically expressed in these cells. EGFP-emission fluorescence spectroscopy revealed that EGCg induced dose-dependent fluorescence changes in EGFP expressing cells, suggesting that EGCg signalling events might trigger proximity changes of EGFP expressed in these cells. Proteomics studies showed that EGFP formed complexes with the 67 kD laminin receptor, caveolin-1 and -3, β-actin, myosin 9, vimentin in EGFP expressing cells. Using in vitro oxidative stress and in vivo myocardial ischemia models, we also demonstrated the involvement of caveolin in EGCg-mediated cardioprotection. In addition, EGCg-mediated caveolin-1 activation was found to be modulated by Akt/GSK-3β signalling in H(2)O(2)-induced H9c2 cell injury. CONCLUSIONS: Our data suggest that caveolin serves as a membrane raft that may help mediate cardioprotective EGCg transmembrane signalling. |
format | Online Article Text |
id | pubmed-3871020 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-38710202013-12-27 Epigallocatechin-3-gallate-mediated cardioprotection by Akt/GSK-3β/caveolin signalling in H9c2 rat cardiomyoblasts Hsieh, Shih-Ron Hsu, Chen-Sen Lu, Chen-Hua Chen, Wei-Cheng Chiu, Chun-Hwei Liou, Ying-Ming J Biomed Sci Research BACKGROUND: Epigallocatechin-3-gallate (EGCg) with its potent anti-oxidative capabilities is known for its beneficial effects ameliorating oxidative injury to cardiac cells. Although studies have provided convincing evidence to support the cardioprotective effects of EGCg, it remains unclear whether EGCg affect trans-membrane signalling in cardiac cells. Here, we have demonstrated the potential mechanism for cardioprotection of EGCg against H(2)O(2)-induced oxidative stress in H9c2 cardiomyoblasts. RESULTS: Exposing H9c2 cells to H(2)O(2) suppressed cell viability and altered the expression of adherens and gap junction proteins with increased levels of intracellular reactive oxygen species and cytosolic Ca(2+). These detrimental effects were attenuated by pre-treating cells with EGCg for 30 min. EGCg also attenuated H(2)O(2)-mediated cell cycle arrest at the G1-S phase through the glycogen synthase kinase-3β (GSK-3β)/β-catenin/cyclin D1 signalling pathway. To determine how EGCg targets H9c2 cells, enhanced green fluorescence protein (EGFP) was ectopically expressed in these cells. EGFP-emission fluorescence spectroscopy revealed that EGCg induced dose-dependent fluorescence changes in EGFP expressing cells, suggesting that EGCg signalling events might trigger proximity changes of EGFP expressed in these cells. Proteomics studies showed that EGFP formed complexes with the 67 kD laminin receptor, caveolin-1 and -3, β-actin, myosin 9, vimentin in EGFP expressing cells. Using in vitro oxidative stress and in vivo myocardial ischemia models, we also demonstrated the involvement of caveolin in EGCg-mediated cardioprotection. In addition, EGCg-mediated caveolin-1 activation was found to be modulated by Akt/GSK-3β signalling in H(2)O(2)-induced H9c2 cell injury. CONCLUSIONS: Our data suggest that caveolin serves as a membrane raft that may help mediate cardioprotective EGCg transmembrane signalling. BioMed Central 2013-11-19 /pmc/articles/PMC3871020/ /pubmed/24251870 http://dx.doi.org/10.1186/1423-0127-20-86 Text en Copyright © 2013 Hsieh et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Hsieh, Shih-Ron Hsu, Chen-Sen Lu, Chen-Hua Chen, Wei-Cheng Chiu, Chun-Hwei Liou, Ying-Ming Epigallocatechin-3-gallate-mediated cardioprotection by Akt/GSK-3β/caveolin signalling in H9c2 rat cardiomyoblasts |
title | Epigallocatechin-3-gallate-mediated cardioprotection by Akt/GSK-3β/caveolin signalling in H9c2 rat cardiomyoblasts |
title_full | Epigallocatechin-3-gallate-mediated cardioprotection by Akt/GSK-3β/caveolin signalling in H9c2 rat cardiomyoblasts |
title_fullStr | Epigallocatechin-3-gallate-mediated cardioprotection by Akt/GSK-3β/caveolin signalling in H9c2 rat cardiomyoblasts |
title_full_unstemmed | Epigallocatechin-3-gallate-mediated cardioprotection by Akt/GSK-3β/caveolin signalling in H9c2 rat cardiomyoblasts |
title_short | Epigallocatechin-3-gallate-mediated cardioprotection by Akt/GSK-3β/caveolin signalling in H9c2 rat cardiomyoblasts |
title_sort | epigallocatechin-3-gallate-mediated cardioprotection by akt/gsk-3β/caveolin signalling in h9c2 rat cardiomyoblasts |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3871020/ https://www.ncbi.nlm.nih.gov/pubmed/24251870 http://dx.doi.org/10.1186/1423-0127-20-86 |
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