摘要
目的探讨miR-214对过氧化氢诱导的心肌细胞凋亡中的影响。方法以心肌细胞H9C2为研究对象,心肌细胞分为4组,依次为正常组、过氧化氢组、miR-214组、NC组,其中miR-214组、NC组细胞分别在转染miR-214模拟物和阴性对照序列后用过氧化氢处理,过氧化氢组用未转染的心肌细胞经过氧化氢处理,正常组用未转染的心肌细胞正常培养。RT-PCR检测细胞中miR-214表达,MTT检测细胞增殖,流式细胞术检测细胞凋亡,Western blot检测细胞中活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、信号转导与转录因子3(STAT3)、磷酸化信号转导与转录因子3(p-STAT3)水平。结果过氧化氢组细胞中miR-214、p-STAT3表达水平和细胞存活率与正常组相比明显下降,而细胞凋亡率及细胞中Cleaved Caspase-3表达水平与正常组相比明显升高(P<0.01)。miR-214组细胞中miR-214、p-STAT3表达水平和细胞存活率与过氧化氢组相比明显升高,而细胞凋亡率及细胞中Cleaved Caspase-3表达水平与过氧化氢组相比明显降低(P<0.01)。结论过氧化氢能够诱导心肌细胞凋亡,miR-214能抑制过氧化氢诱导的心肌细胞凋亡,STAT3信号通路可能是其作用机制之一。
Objective To investigate the role of miR -214 in the apoptosis of myocardial cells induced by hydrogen peroxide. Methods H9C2 was used as the research object, the myocardial cells were divided into 4 groups： the normal group, the hydrogen peroxide group, the miR -214 group and the NC group, the miR -214 group and NC group cells were transfected with miR -214 and negative control sequences, then hy- drogen peroxide treatment, Hydrogen peroxide group was treated with hydrogen peroxide, the normal group was cultured with the normal culture of the non transfected cardiomyocytes. The miR -214 expression in RT- PCR cells, cell proliferation was detected by MTT, cell apoptosis was de- tected by flow cytometry. Western blot was used to detect Cleaved Caspase - 3, STAT3 and p - STAT3 levels. Results The expression levels of miR - 214, p - STAT3 and cell survival rate in hydrogen peroxide group were significantly lower than those in normal group, the apoptosis rate and the expression of Cleaved Caspase -3 in the cells were significantly higher than those in the normal group （ P〈0.01 ）. The expression of miR - 214, p -STAT3 and cell survival rate in group miR -214 were significantly higher than those in the control group, the apoptosis rate and the ex-pression of Cleaved Caspase - 3 in the cells were significantly lower than those in the control group （ P 〈 0.01 ）. Conclusion Hydrogen peroxide can induce cardiomyocyte apoptosis, miR -214 can inhibit the apoptosis of myocardial cells induced by hydrogen peroxide, STAT3 signaling path- way may be one of its mechanisms.
出处
《临床和实验医学杂志》
2017年第17期1692-1695,共4页
Journal of Clinical and Experimental Medicine
