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目的探讨microRNA-29a(miR-29a)对大鼠心肌细胞凋亡的调控作用。方法体外培养新生SD大鼠心肌细胞,合成人miR-29a的拟似物(mimic)。用Lipofectamine RNAiMAX转染miR-29a的mimic进入心肌细胞,转染48 h后用荧光定量PCR方法检测心肌细胞miR-29a的表达变化,流式细胞仪检测细胞凋亡水平变化,Westernblot法检测凋亡相关蛋白Caspase-3和Caspase-9前体的表达变化。结果心肌细胞转染miR-29a的mimic 48 h后,心肌细胞中miR-29a的表达水平较对照组明显升高(P<0.05);心肌细胞的凋亡水平也明显升高,凋亡相关蛋白Caspase-3和Caspase-9前体的含量则明显下降(P<0.05)。结论在大鼠心肌细胞中过表达miR-29a能促进心肌细胞凋亡,其机制可能是通过Caspase-3和Caspase-9途径起作用。
Objective To investigate the regulatory effect of microRNA-29a (miR-29a) on cardiomyocyte apoptosis in rats. Methods Cardiomyocytes from neonatal SD rats were cultured in vitro and the mimic of human miR-29a was synthesized. The mimic of miR-29a was transfected into cardiomyocytes with Lipofectamine RNAiMAX. The expression of miR-29a in cardiomyocytes was detected by fluorescence quantitative PCR 48 h after transfection. The apoptosis was detected by flow cytometry and the apoptosis was detected by Western blot Changes of Caspase-3 and Caspase-9 Precursor Protein Expression. Results The expression of miR-29a in cardiomyocytes was significantly increased in myocardial cells transfected with mimic miR-29a for 48 h (P <0.05), and the apoptosis of cardiomyocytes was also significantly increased. The expression of apoptosis-related proteins Caspase-3 and Caspase-9 precursor levels were significantly decreased (P <0.05). Conclusion Overexpression of miR-29a in rat cardiomyocytes can promote cardiomyocyte apoptosis, and its mechanism may be through the Caspase-3 and Caspase-9 pathways.