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- 2016
microRNA-499靶控的PI3K/Akt通路在大鼠缺血-再灌注心肌损伤中的作用*
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Abstract:
目的:探讨microRNA-499在大鼠心肌缺血-再灌注损伤(MIRI)中的作用及其与PI3K/Akt通路的关系。方法:将30只SD大鼠随机分为假手术组、模型组和干预组,每组10只。假手术组仅在冠状动脉的左前降支下穿线但并不结扎,模型组结扎冠状动脉左前降支以建立大鼠MIRI模型(缺血30 min,再灌注120 min),干预组于建立MIRI模型前48 h注射microRNA-499激动剂。采用荧光定量RT-PCR法检测大鼠心肌组织中miRNA-499及PI3K mRNA表达量,Western blot法检测Akt磷酸化活性,采用TTC染色法检测心肌梗死面积。结果:与假手术组相比,模型组miRNA-499及PI3K mRNA表达量减少,Akt磷酸化活性明显降低,心肌梗死面积明显增加(P＜0.05)。与模型组比较,干预组miRNA-499及PI3K mRNA表达量、Akt磷酸化活性明显升高,心肌梗死面积明显减少(P＜0.05)。心肌组织中PI3K mRNA与microRNA-499的表达量呈正相关(P＜0.05)。结论:microRNA-499可能通过激活PI3K/Akt信号通路,减轻大鼠MIRI。
Aim: To investigate the effect of miRNA-499 on myocardial ischemia-reperfusion injury(MIRI)of rats and its relation with PI3K/Akt signaling pathway.Methods: Thirty SD rats were randomly assigned into sham-operation group, model group, and precondition group with 10 rats in each group. In the sham-operation group, the thread was only threaded under left anterior descending coronary artery(LAD)but not ligated; in model group, MIRI models were built by ligating LAD with 30 min ischemia and 120 min reperfusion; in the precondition group, rats were injected miRNA-499 agomir 48 h before MIRI operation. Real Time RT-PCR was adopted to detect the expressions of miRNA-499 and PI3K mRNA, Western blot was adopted to detect Akt phosphorylation level, and the myocardial infarction size were measured using TTC staining.Results: Compared with sham-operation group, the expressions of miRNA-499 and PI3K mRNA, and the Akt phosphorylation level in model group were significantly lower,and the infarction size were larger(P＜0.05). Compared with those in MIRI group, the expressions of miRNA-499 and PI3K mRNA, and the Akt phosphorylation level in precondition group were obviously higher,while the infarction size was smaller(P＜0.05). The expression of microRNA-499 was positively related to that of PI3K mRNA(P＜0.05)。Conclusion: microRNA-499 could reduce MIRI via activating the PI3K/Akt signaling pathway
