miRNA-29c对大鼠心肌细胞增殖和凋亡的影响及机制研究

Effect of miRNA-29c on proliferation and apoptosis of rat cardiomyocytes and its mechanism

目的:探讨miRNA-29c对大鼠心肌细胞增殖和凋亡的影响及可能机制。方法:将大鼠H9C2心肌细胞分为空白对照(control)组、miRNA-29c模拟物转染(LV-miR-29c-mimic)组、阴性转染(LV-NC)组。实时荧光定量聚合酶链式反应(RT-qPCR)检测各组miRNA-29c、VEGF的mRNA表达情况,免疫荧光染色法检测各组细胞VEGF、Bax、Bcl-2、caspase3荧光表达,细胞计数试剂盒(CCK-8)检测各组细胞增殖情况,蛋白免疫印迹法(Western blotting)检测各组VEGF、Bax、Bcl-2、caspase3的蛋白表达情况,流式细胞仪检测各组细胞凋亡率。结果:与control组和LV-NC组相比,LV-miR-29c-mimic组H9C2大鼠心肌细胞miRNA-29c表达水平升高,细胞增殖速率、VEGF mRNA及其蛋白、Bcl-2的蛋白表达降低,细胞凋亡率、Bax、caspase3的蛋白表达则增高(均P<0.05)。结论:miRNA-29c可能通过调控细胞内的VEGF表达,影响大鼠心肌细胞的增殖和凋亡。

Objective:To investigate the effect of miRNA-29c on the proliferation and apoptosis of rat cardiomyocytes and its possible mechanism.Methods:Rat H9C2 cardiomyocytes were divided into blank control group,miRNA-29c mimic transfected(LV-miR-29c-mimic)group,and negative transfected(LV-NC)group.Real-time fluorescence quantitative PCR(RT-qPCR)was performed to detect the expression of miRNA-29c and VEGF.The fluorescence expression of VEGF,Bax,Bcl-2 and caspase3 was measued by immunofluorescence staining.The cell proliferation was detected by CCK-8 assay.The protein expression of VEGF,Bax,Bcl-2 and caspase3 was assessed by western blotting.The apoptosis rate was determined by flow cytometry.Results:Compared with the blank control group and LV-NC group,the expression of miRNA-29c in H9C2 rat cardiomyocytes was increased,cell proliferation,the mRNA and protein levels of VEGF and the protein level of Bcl-2 were decreased,while the apoptosis rate and the protein expression of Bax and caspase3 were increased in the LV-miR-29c-mimic group(P<0.05).Conclusion:miRNA-29c may affect the proliferation and apoptosis of rat cardiomyocytes by regulating intracellular VEGF expression.