基础医学与临床 ›› 2019, Vol. 39 ›› Issue (5): 677-681.

• 研究论文 • 上一篇    下一篇

miR-138在乙醇诱导的大鼠心肌细胞系H9C2氧化应激中表达增加

王圣凯,胡苏琴,吴玲,吴杭菲,郑靖宇,蒋自培,李剑敏   

  1. 温州医科大学附属第一医院
  • 收稿日期:2018-12-18 修回日期:2019-03-18 出版日期:2019-05-05 发布日期:2019-04-24
  • 通讯作者: 李剑敏 E-mail:wzyxyljmin@163.com
  • 基金资助:
    浙江省自然科学基金;浙江省自然科学基金

miR-138 expression is increased in rat cardiomyocyte cell line H9C2 with myocardial oxidative stress

  • Received:2018-12-18 Revised:2019-03-18 Online:2019-05-05 Published:2019-04-24
  • Contact: Min JianLI E-mail:wzyxyljmin@163.com

摘要: 目的 观察miR-138对乙醇诱导的大鼠心肌细胞系H9C2的Sirt1表达及其下游氧化应激和线粒体凋亡相关蛋白表达的影响。方法 将心肌细胞分为对照组(control组)、乙醇组(E组)、miR-138抑制剂组(I组)。分别用不同浓度的含乙醇的细胞培养液培养,MTT法检测H9C2细胞的活力,选择细胞活力接近80%的200 mmol/L乙醇浓度为最佳干预浓度。用Lipo2000,将miR-138抑制剂转染I组H9C2心肌细胞,孵育12 h后,分别用羟胺法测定心肌细胞中总超氧化物歧化酶(T-SOD)活力、TBA法测定心肌细胞丙二醛(MDA)含量、用RT-qPCR检测心肌细胞内miR-138与Sirt1 mRNA的表达、用Western blot测定心肌细胞内Sirt1、Bcl-2、Bax 和caspase-3蛋白的表达。结果 1)乙醇可以抑制心肌细胞的活力。2)与对照组相比,乙醇组心肌细胞中miR-138 mRNA的表达及MDA的含量、Bax和caspase-3的蛋白表达均明显升高(P<0.05),T-SOD的活力、Sirt1 mRNA的表达及Sirt1和Bcl-2蛋白的表达均明显降低(P<0.05);与乙醇组相比,miR-138抑制剂组心肌细胞中miR-138 mRNA的表达及MDA的含量及Bax、caspase-3的蛋白表达均明显降低(P<0.05),T-SOD的活力、Sirt1 mRNA的表达、Sirt1和Bcl-2蛋白的表达和均明显升高(P<0.05)。结论 miR-138在乙醇诱导的大鼠H9C2心肌细胞氧化应激中表达增加,Sirt1可能是miR-138的作用靶点。

关键词: 乙醇性心肌病, miR-138, Sirt1, 氧化应激, 凋亡

Abstract: Objective To observe the changes of miR-138 on Sirt1 expression in ethanol induced Rat H9C2 myocardial cell lines and its downstream oxidative stress and mitochondrial apoptosis-related protein expression. Methods Cells were divided into control group, ethanol group (E group) and miR-138 inhibitor group (I group), every groups were cultured with different concentrations of alcohol culture media for 24h, cell viability was detected by MTT assay and 200mmol/L alcohol culture media was selected as the optimal concentration for following experiment. Group I was cultured with miR-138 inhibitor transfection media. After 12-hour-incubation, total superoxide dismutase (T-SOD) activity was analysed by hydroxylamine method and the content of malondialdehyde (MDA) was detected by thiobarbituric acid method. The expression of mRNA of miR-138 and Sirt1 were detected by RT-qPCR. The expression of Sirt1, Bax, Bcl-2 and caspase-3 protein was detected by Western blot. Results 1.Cell viability could be suppressed by alcohol. 2.Compared with control group, the content of MDA, the expression of miR-138 and both the protein expressions of Bax and caspase-3 were increased significantly (P<0.05) in ethanol group, the T-SOD activity, the mRNA expression of Sirt1, protein expressions of Sirt1 and Bcl-2 decreased significantly (P<0.05) in ethanol group. Compared with ethanol group, the content of MDA, the expression of miR-138 and both the protein expressions of Bax and caspase-3 decreased significantly (P<0.05) in miR-138 inhibitor group, the T-SOD activity, the mRNA expression of Sirt1, protein expressions of Sirt1 and Bcl-2 increased significantly (P<0.05) in miR-138 inhibitor group. Conclusions miR-138 increases oxidative stress of rat H9C2 cardiomyocytes induced by ethanol, Sirt1 may be the target of miR-138.

Key words: alcoholic cardiomyopathy, miR-138, silent information regulator 1, oxidative stress, apoptosis

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