过表达miR-29a对大鼠心肌细胞凋亡的影响
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Overexpression of MicroRNA29a Regulates Cells Apoptosis in Rat Cardiomyocytes
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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,Western blot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。

    Abstract:

    AimTo investigate the role of microRNA-29a (miR-29a) on apoptosis in rat cardiomyocytes.MethodsIn our study, newborn rats cardiomyocytes was isolated, cells were transfected with miR-29a mimic by lipofectamine RNAiMAX, and then real time RT-PCR was used to measure the level of miR-29a, flow cytometry (FCM) was used to detect the cells apoptosis, and western blot was used to detect the protein expressive level of Caspase-3 and Caspase-9.ResultsMiR-29a mimics could upregulate the level of miR-29a in rat cardiomyocytes after 48h of transfecting, accompanied with the increase of cells apoptosis rate, and descent of the protein level of caspase-3 and caspase-9 in cardiomyocytes when compared to the control group(P<0.05).ConclusionOur study demonstrated that the overexpression of miR-29a may regulate cells apoptosis of rat cardiomyocytes, and the mechanisms among it may be concerned with Caspase-3 and Caspase-9 pathway.

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张振辉,黎 佼,刘本荣,李爱群,刘世明,.过表达miR-29a对大鼠心肌细胞凋亡的影响[J].中国动脉硬化杂志,2012,20(12):1079~1082.

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  • 收稿日期:2012-07-16
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