大黄酚对缺氧诱导的心肌细胞损伤保护作用及机制

Protective effect of chrysophanol on hypoxia-induced cardiomyocyte injury and its mechanism

目的探讨大黄酚对缺氧诱导的心肌细胞H9c2损伤的保护作用及其机制。方法将体外培养的H9c2细胞分为对照组(正常培养)、缺氧组(缺氧处理)和1、10、20μmol/L大黄酚处理组(1、10和20μmol/L大黄酚处理后,给予缺氧处理)。采用LDH试剂盒检测细胞上清液中LDH释放量,MTT法检测细胞活力,流式细胞仪检测细胞凋亡,Western blot检测细胞中Bcl-2、Bax和Cleaved caspase-3蛋白水平。结果与对照组相比,缺氧组细胞上清液中LDH释放量明显增多,细胞存活率和细胞中Bcl-2/Bax水平明显降低,细胞凋亡率和Cleaved caspase-3蛋白的表达水平明显升高(P<0.05);与缺氧组相比,1μmol/L大黄酚处理对细胞无明显影响(P>0.05),但10、20μmol/L大黄酚处理后缺氧引起的上述变化明显得到改善(P<0.05),且20μmol/L大黄酚处理组的作用效果明显高于10μmol/L大黄酚处理组(P<0.05)。结论大黄酚可通过拮抗缺氧诱导的心肌细胞凋亡而保护心肌细胞损伤。

Objective To investigate the protective effect of chrysophanol on hypoxia-induced H9c2 injury in cardiomyocytes and its mechanism.Methods H9c2 cells cultured in vitro were divided into control group(normal culture),hypoxia group(hypoxia treatment)and 1,10,20μmol/L chrysophanol treatment group(1,10,20μmol/L chrysophanol treatment,hypoxia treatment).LDH release in cell supernatant was detected by LDH kit,cell viability was detected by MTT assay,apoptosis was detected by flow cytometry,and the expression of bcl 2,Bax and cleaved caspase 3 protein was detected by Western blot.Results Compared with the control group,LDH release in the supernatant of hypoxic group significantly increased,cell survival rate and Bcl-2/Bax level significantly decreased,apoptotic rate and expression of Cleaved caspase-3 protein significantly increased(P<0.05).Compared with the hypoxic group,1μmol/L chrysophanol treatment had no significant effect on cells(P>0.05).However,the above changes induced by hypoxia after 10 and 20μmol/L chrysophanol treatment were significantly improved(P<0.05),and the effect of 20 micromol/L chrysophanol treatment group was significantly better than that of 10μmol/L chrysophanol treatment group(P<0.05).Conclusion Chrysophanol protects cardiomyocyte injury by antagonizing hypoxia-induced cardiomyocyte apoptosis.