天津医药

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miRNA-21与JAK2-STAT3通路在心肌缺血后处理保护心肌中的相互影响*

薛才广1,梁德刚2,魏民新1,田轶魁2   

  1. 1. 天津医科大学总医院心血管外科
    2. 天津医科大学总医院
  • 收稿日期:2012-09-19 修回日期:2012-12-24 出版日期:2013-05-15 发布日期:2013-05-15
  • 通讯作者: 田轶魁

Crosstalk between miRNA-21 and JAK2-STAT3 pathway in protective effect of myocardial ischemic postconditioning

XUE CAI GUANG,LIANG DE GANG,WEI MIN XIN,QIAN YI KUI   

  1. Department of Cardiovascular Surgery, General Hospital of Tianjin Medical University
  • Received:2012-09-19 Revised:2012-12-24 Published:2013-05-15 Online:2013-05-15
  • Contact: QIAN YI KUI

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摘要： 【摘要】目的 观察miRNA-21在缺血后处理心肌保护机制中的作用，并探讨其与JAK2-STAT3通路在心肌缺血后处理保护心肌机制中的相互影响。方法 将健康雄性成年Wistar大鼠随机分为假手术组（Sham组）、缺血再灌注损伤组(I/R组)、缺血后处理组（PostC组）、缺血后处理+miRNA-21抑制物处理组（PostC+antagomiR-21组）和缺血后处理+AG490组（PostC+AG490组）。原位末端脱氧核苷酸转移酶标记（TUNEL）法检测心肌细胞凋亡指数；Western blot法检测总STAT3（t-STAT3）及磷酸化STAT3（p-STAT3）蛋白水平；荧光实时定量（qRT）-PCR检测心肌组织miRNA-21表达水平。结果  与I/R组相比，PostC组再灌注后心肌细胞凋亡指数显著减少，miRNA-21表达水平及STAT3磷酸化水平显著上调。使用antagomiR-21可显著削弱PostC心肌保护作用。PostC+antagomiR-21组与PostC组相比，STAT3的磷酸化水平显著降低。PostC+AG490组与PostC组比较，STAT3的磷酸化水平及miRNA-21表达均显著降低。结论 miRNA-21在PostC心肌保护机制中发挥调控作用。在后处理适应性机制中STAT3与miRNA-21可能存在某种回路调控机制。

关键词: 心肌再灌注损伤, 微RNAs, 原位缺口末端标记, 细胞凋亡, STAT3转录因子, 大鼠, Wistar, 缺血后处理

Abstract: [Abstract] Objective  To investigate the protective effect of miRNA-21 in myocardial ischemic postconditioning, and to explore the interaction effects in the mechanism of myocardial protection with JAK2-STAT3 pathway in ischemic postconditioning. Methods   The healthy adult male Wistar rats (body weight 240~280g) were randomly divided into five groups:Sham group, schemia-reperfusion injury group (I/R group), Ischemic postconditioning group (PostC group), schemic postconditioning + antagomiR-21 treatment group (PostC + antagomiR-21 group), and Ischemic postconditioning + AG490 group (PostC + AG490 group). In situ terminal deoxynucleotidyl transferase labeling (TUNEL) staining was used to analysis myocardial apoptosis index; Western-blot was used to detect total STAT3 (t-stat3) and phospho-STAT3 (p-stat3) protein levels.miRNA-21 levels were detected by quantitative reverse transcriptase–polymerase chain reaction (qRT-PCR). Results   Compared with the I / R group after reperfusion,ischemic postconditioning significantly reduced the apoptosis index, dramatically increased the expression levels of miRNA-21 and markedly up-regulated the stat3 phosphorylation levels. AntagomiR-21 significantly weaken the protective effect of ischemic postconditioning. PostC + antagomiR-21 group compared with PostC, STAT3 phosphorylation levels were observably lower. Compared with postC group, STAT3 phosphorylation levels and miRNA-21 were significantly lower in PostC+AG490 group. Conclusion   miRNA-21 play a regulatory role in myocardial ischemic postconditioning. STAT3 and miRNA-21 in post-processing adaptive mechanism there may be some kind of loop regulatory mechanism. But the reciprocal of the detailed regulation mechanism is unclear, pending further study.

Key words: myocardial reperfusion injury, microRNAs, in situ nick-end labeling, cell cycles, STAT3 transcription factor, rats, Wistar, 缺血后处理