天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (10): 1217-1220.doi: 10.11958/20160435

• 实验研究 • 上一篇    下一篇

miR-34a改变自噬通路AMPK/mTOR的活性在心肌细胞肥大中的作用

黄炯华,林育辉,戴文军,伍金雷,谢文杰,陈永权   

  1. 广州医科大学附属第三医院心血管内科（邮编 510150）
  • 收稿日期:2016-05-17 修回日期:2016-06-22 出版日期:2016-10-15 发布日期:2016-10-21
  • 通讯作者: 黄炯华 E-mail:mdhjh2014@126.com
  • 作者简介:黄炯华（1984）， 男， 博士， 主要从事心肌肥厚的作用机制及临床相关研究
  • 基金资助:
    广东省自然科学基金项目（2015A030310068）；广州医科大学博士启动基金项目（2014C37）

The role of miR-34a in AngⅡ induced myocardial hypertrophy via alteration of AMPK/mTOR signal pathway

HUANG Jionghua, LIN Yuhui, DAI Wenjun, WU Jinlei, XIE Wenjie, CHEN Yongquan   

  1. Department of Cardiology, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China
  • Received:2016-05-17 Revised:2016-06-22 Published:2016-10-15 Online:2016-10-21
  • Contact: HUANG Jionghua E-mail:mdhjh2014@126.com

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摘要： 目的 研究 miR-34a 与自噬通路磷酸腺苷蛋白激酶（AMPK）/哺乳动物雷帕霉素靶点（mTOR）在血管紧张素（Ang）Ⅱ 诱导的原代大鼠心肌细胞肥大中的作用。 方法 体外培养原代大鼠心肌细胞， 分成 3 组： 阴性对照慢病毒（NC）组、AngⅡ 刺激+阴性对照慢病毒（AngⅡ +NC）组、AngⅡ 刺激+miR-34a 过表达慢病毒（AngⅡ +miR-34a）组。激光共聚焦检测心肌细胞肥大变化； 利用荧光定量聚合酶链反应（qRT-PCR）检测 miR-34a、心房钠尿肽（ANP）和 β- 肌球蛋白重链（β-MHC）表达； 利用蛋白印迹（Western blot）测定 AMPK/mTOR 信号通路的活性变化。 结果 与 NC 组比较， AngⅡ +NC 组心肌细胞表面积明显增大（P＜ 0.05）； 与 AngⅡ +NC 组比较， AngⅡ +miR-34a 组心肌细胞表面积减少（P＜ 0.05）。 与 NC 组比较， AngⅡ +NC 组的 miR-34a 表达下调， ANP 和 β-MHC 表达上调（均 P＜ 0.05）； 而与 AngⅡ +NC 组比较， AngⅡ +miR-34a 组的 miR-34a 表达上调， ANP 和 β-MHC 表达下调（均 P＜ 0.05）。 与 NC 组比较， AngⅡ +NC 组 p-AMPK/AMPK 增加， p-mTOR/mTOR 减少（均 P＜ 0.05）； 与 AngⅡ +NC 组比较， AngⅡ +miR-34a 组 p-AMPK/AMPK 减少， p-mTOR/mTOR 增加（均 P＜ 0.05）。 结论 miR-34a 能抑制 AngⅡ 诱导的心肌细胞肥大， 其作用部分是通过改变 AMPK/mTOR 的活性来实现的。

关键词: 肌细胞, 心脏, 心肌病, 肥厚性, 血管紧张素Ⅱ , 疾病模型, 动物, 自噬通路磷酸腺苷蛋白激酶/哺乳动物雷帕霉素靶点, miR-34a

Abstract: Objective To explore the effects of miR- 34a and the alteration of AMPK/mTOR signal pathway on angiotensin (Ang)Ⅱ -stimulated cardiomyocytes hypertrophy. Methods Neonatal rat cardiomyocytes were cultrued in vitro and were divided into 3 groups: negative control for lentivirus carrying miR-34a group (NC), AngⅡ plus lentivirus carrying negative control group (AngⅡ +NC) and AngⅡ plus lentivirus carrying miR-34a group (AngⅡ +miR-34a). The relative cell area was detected by confocal microscopy. The expression of miR-34a and hypertrophy-related genes (ANP and β-MHC) were analyzed by real time PCR. The AMPK/mTOR signal pathway was measured by Western blot assay. Results Compared to NC group, the relative cell area was increased in AngⅡ +NC group (P＜ 0.05). But compared with AngⅡ +NC group, the relative cell area was decreased in AngII+miR- 34a group (P＜ 0.05). Moreover, compared with NC group, the expression of miR- 34a was decreased, and the expression of hyperthophy- related genes(ANP and β- MHC) was upregulated in AngⅡ +NC group. However, the expression of miR- 34a was decreased, and the expression of hyperthophyrelated genes (ANP and β-MHC) was down-regulated (P＜ 0.05). Finally, compared to NC group, the ratio of phosphopAMPK/AMPK was significantly induced in AngII + NC group, but the ratio of phosphop-mTOR/mTOR was significantly decreased (P＜ 0.05). However, compared to Ang Ⅱ + NC group, the ratio of phosphop- AMPK/AMPK was significantly decreased in AngII + miR- 34a group, but the ratio of phosphop- mTOR/mTOR was significantly increased (P＜ 0.05). Conclusion miR-34a is able to inhibit myocardial hypertrophy of neonatal rat cardiomyocytes, and its mechanism is partly carried out by the alteration of the signal pathway of AMPK/mTOR.

Key words: myocytes, cardiac, cardiomyopathy, hypertrophic, angiotensinⅡ , disease models, animal, AMPK/ mTOR, miR-34a