Wang Lifeng, Lu Yingjie, Liu Xian, Wang Xiaoyun
Department of Cardiology, The Fourth Clinical Hospital of Harbin Medical University, No. 37 Yiyuan Road, Nangang District, Harbin, 150001, Heilongjiang, China.
Mol Biol Rep. 2017 Apr;44(2):219-226. doi: 10.1007/s11033-017-4098-z. Epub 2017 Mar 9.
Reducing reperfusion period myocardial cell damage is efficient to reduce myocardial ischemia-reperfusion injury. Ghrelin can increase myocardial contractility, improve heart failure caused by myocardial infarction. This study aimed to investigate the protective effect of Ghrelin on myocardial hypoxia/reoxygenation (H/R) injury of neonatal rat cardiomyocytes (NRCMs) and to explore the mechanisms. We isolated the NRCMs, established myocardial H/R model, blocked growth hormone secretagogue receptor (GHSR) by siRNA technique, examined cell activity by MTT and LDH assay, detected apoptosis by Hoechst 33258 staining and flow cytometry and determined the expression levels of apoptosis related proteins and signaling pathway proteins by western blot. We found that Ghrelin can significantly improve cell activity and decrease apoptosis after H/R, however this effect was abolished by GHSR-siRNA. In addition, we found that Ghrelin can significantly increase the expression of Bcl-2 but inhibit the level of Bax and caspase-3. Further mechanism study found that the phosphorylation level of signaling pathway protein Akt and mTOR in Ghrelin treated group were significantly higher than that in other groups. In conclusion, Ghrelin can reduce the H/R damage on NRCMs and inhibit the apoptosis by activating Akt-mTOR signaling pathway.
减少再灌注期心肌细胞损伤对于减轻心肌缺血-再灌注损伤是有效的。胃饥饿素可增强心肌收缩力,改善心肌梗死所致的心力衰竭。本研究旨在探讨胃饥饿素对新生大鼠心肌细胞(NRCMs)缺氧/复氧(H/R)损伤的保护作用并探究其机制。我们分离了NRCMs,建立心肌H/R模型,采用小干扰RNA(siRNA)技术阻断生长激素促分泌素受体(GHSR),通过MTT法和乳酸脱氢酶(LDH)检测法检测细胞活性,采用Hoechst 33258染色和流式细胞术检测细胞凋亡,并通过蛋白质免疫印迹法测定凋亡相关蛋白和信号通路蛋白的表达水平。我们发现胃饥饿素可显著改善H/R后细胞活性并减少细胞凋亡,然而这种作用被GHSR-siRNA消除。此外,我们发现胃饥饿素可显著增加Bcl-2的表达,但抑制Bax和半胱天冬酶-3的水平。进一步的机制研究发现,胃饥饿素处理组中信号通路蛋白Akt和哺乳动物雷帕霉素靶蛋白(mTOR)的磷酸化水平显著高于其他组。总之,胃饥饿素可减轻NRCMs的H/R损伤并通过激活Akt-mTOR信号通路抑制细胞凋亡。