Gastrodin inhibits apoptosis of H9c2 cardiomyocytes through protein kinase B/p38mitogen actived protein kinase signaling pathway

doi:10.16098/j.issn.0529-1356.2019.01.008

Abstract

Abstract:

Objective To investigate the effect and precise molecular mechanisms for explaining how gastrodin suppresses the apoptosis in the serum deprivation/reperfusion-induced H9c2 cardiomyocytes. Methods Rat H9c2 cardiomyocytes were cultured and treated differently in vitroand randomly divided into following groups, including the control group, the serum deprivation-stimulated (0.5-6 hours) groups, the serum deprivation/reperfusion(2/4 hours) group, gastrodin (10, 20 and 40 μmol/L) treatment groups, wortmannin (1 μmol/L) and gastrodin(20 μmol/L) co-treatment group, wortmannin (1 μmol/L) treatment group. The variation of protein expression levels of p-p38 mitogen actived protein kinase(MAPK), p-Akt and apoptosis related cleaved caspase-3, Bcl-2 and Bax were analyzed by Western blotting.H9c2 cardiomyocytes were analyzed by flow cytometry.The level of RNA expression levels of apoptosis related Caspase-3,Bcl-2 and Bax were analyzed by Real-time PCR assay. The expression level of p-Akt was assayed by double immunofluorescence labeling. Results The results showed that serum deprivation (0.5-6 hours) and serum deprivation/reperfusion (2/4 hours) induced apoptosis increasing; Using Western blotting and Real-time PCR assay, we found that treatment with gastrodin significantly alleviated cleaved Caspase-3 activation and increased the ratio of Bcl-2/Bax in a concentration-dependent manner after serum deprivation and serum deprivation/reperfusion exposure. The same result has been showed by flow cytometer. Serum deprivation and serum deprivation/reperfusion inhibited the Akt/p38MAPK signaling pathway; After treatment with gastrodin in a concentration-dependent manner induced the expression of p-Akt and decreased the expression of p-p38MAPK. The regulation effects of gastrodin were reversed by the Akt specific inhibitor wortmannin. Conclusion In conclusion , the present study demonstrated that gastrodin exerted protective effects against serum deprivation/reperfusion-induced apoptosis in H9c2 cardiomyocytes, at least partly via the Akt/p38MAPK pathway.

Key words: Gastrodin, Myocardial ischemia/reperfusion injury, p38 mitogen actived protein kinase, Western blotting, Double immunofluorescence labeling, Flow cytometry, Rat

ZHANG Ling YANG Ping JIANG Yong-liang PU Yu-wei XIE Li-qiu SUN Lin LU Di. Gastrodin inhibits apoptosis of H9c2 cardiomyocytes through protein kinase B/p38mitogen actived protein kinase signaling pathway[J]. Acta Anatomica Sinica, 2019, 50(1): 40-48.

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