天津医药

• 细胞与分子生物学 •    下一篇

下调microRNA-1对H2O2诱导的心肌细胞损伤的保护作用

郑君毅   

  1. 天津市胸科医院
  • 收稿日期:2013-10-22 修回日期:2014-03-27 出版日期:2014-08-15 发布日期:2014-08-15
  • 通讯作者: 郑君毅

Downregulation MicroRNA-1Can Protect H2O2 Injured Cardiomyocytes

  • Received:2013-10-22 Revised:2014-03-27 Published:2014-08-15 Online:2014-08-15

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摘要： 【摘要】目的? ?研究下调microRNA-1在H2O2诱导的心肌细胞损伤中的保护作用及其机制。方法? ?将大鼠H9c2心肌细胞株分为4组，Blank组、NC组、H2O2组和H2O2+AS-miR-1组。Blank组为不做任何处理的空白对照组，NC组细胞转染随机合成的miRNA阴性对照片；H2O2组和H2O2+AS-miR-1组分别为不转染或转染miR-1 inhibitor（AS-miR-1）的H2O2细胞损伤组。采用定量PCR方法检测各组miR-1表达水平，MTT和流式细胞术检测细胞存活率和凋亡情况，利用生物信息学预测miR-1的靶基因，荧光定量PCR和Western Blot的方法检测靶基因Bcl-2的mRNA和蛋白表达情况。结果? ?Blank组和NC组相比较，各个指标均没有统计学差异。H2O2组细胞的miR-1mRNA表达水平显著升高，存活率降低，凋亡增加，Bcl-2的mRNA和蛋白表达水平均明显降低。转染AS-miR-1，可以降低H2O2诱导的心肌细胞损伤，提高细胞存活率、降低细胞凋亡，上升Bcl-2的mRNA和蛋白表达水平。 结论? ?下调miR-1表达可以通过升高凋亡抑制因子Bcl-2的表达水平降低H2O2诱导的心肌细胞凋亡，发挥心肌细胞保护作用。

关键词: miR-1, 细胞凋亡, 转染, 微RNAs, 肌细胞, 心脏, 氧化性应激, 过氧化氢

Abstract: Objective To investigate the protective function and mechanism of microRNA-1 (miR-1) downregulation in H2O2 injured cardiomyocytes. Methods The experiment was divided into 4 groups:Blank, NC, H2O2 and H2O2+AS-miR-1. Blank cells had no treatment, NC cells transfected with random miRNA fragement, H2O2 and H2O2+AS-miR-1 group were defined as H2O2 injured cardiomyocytes without or with transfected antisense miR-1 oligonucleotide（AS-miR-1）. The real time PCR was used to test miR-1 expression, cell vitality and apoptosis were analyzed by MTT and flow cytometry. The target gene of miR-1 was predicted by bioinformatics, and then, the mRNA and protein expression level of Bcl-2 were detected by real time PCR and western blot. Results All index had no significant changes in Blank and NC groups. H2O2 can induce cardiomyocyte injury, increase the miR-1 level and apoptosis rate, reduce cell vitality and Bcl-2 expression level. Transfection AS-miR-1 can decrease cell apoptosis, increase cell vitality and Bcl-2 expression level. Conclusion Downregulation microRNA-1 can protect H2O2 injured cardiomyocytes by increasing anti-apoptosis factor Bcl-2 expression.

Key words: miR-1, cell cycles, transfection, microRNAs, myocytes, cardiac, oxidative stress, 过氧化氢