Epigallocatechin-3-gallate-mediated cardioprotection by Akt/GSK-3β/caveolin signalling in H9c2 rat cardiomyoblasts

Journal of Biomedical Science

Ying-Ming Liou⁴  Chun-Hwei Chiu²  Wei-Cheng Chen¹  Chen-Hua Lu²  Chen-Sen Hsu²  Shih-Ron Hsieh³ 

[1] Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 30013, Taiwan;Department of Life Sciences, National Chung-Hsing University, Taichung 402, Taiwan;Department of Cardiovascular Surgery, Taichung Veterans General Hospital, Taichung 407, Taiwan;Graduate Institute of Basic Medical Science, China Medical University, Taichung 40402, Taiwan

关键词: Oxidative stress;    Caveolin;    EGFP;    H9c2;    Cell cycle;    EGCg;   

Others  :  821765
DOI  :  10.1186/1423-0127-20-86

 received in 2013-08-22, accepted in 2013-11-18,  发布年份 2013

【 摘 要 】

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 H₂O₂-induced oxidative stress in H9c2 cardiomyoblasts.

Results

Exposing H9c2 cells to H₂O₂ suppressed cell viability and altered the expression of adherens and gap junction proteins with increased levels of intracellular reactive oxygen species and cytosolic Ca²⁺. These detrimental effects were attenuated by pre-treating cells with EGCg for 30 min. EGCg also attenuated H₂O₂-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 H₂O₂-induced H9c2 cell injury.

Conclusions

Our data suggest that caveolin serves as a membrane raft that may help mediate cardioprotective EGCg transmembrane signalling.

【 授权许可】

   
2013 Hsieh et al.; licensee BioMed Central Ltd.

【 图 表 】

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