缺血再灌注损伤中miR-126的作用及对心肌细胞凋亡的影响

发布时间：2018-05-25 04:14

本文选题：miR- + 心肌缺血再灌注损伤　；参考：《重庆医科大学学报》2017年09期

【摘要】：目的:通过体内外实验探讨微小RNA(miR)-126在心肌缺血再灌注损伤中的作用及机制。方法:体外部分:心肌细胞H9C2分6组,建立心肌细胞模拟缺血/再灌注(simulated ischemia/reperfusion,SI/R)损伤模型,并分别转染miR-126mimic和miR-126inhibitor。qRT-PCR检测心肌细胞miR-126表达,流式细胞术检测细胞凋亡,Western blot检测caspase-3及其裂解片段的蛋白表达。体内部分:雌性Wistar健康大鼠分5组,建立缺血/再灌注损伤(ischemia/reperfusion,I/R)模型,并分别转染含有GFP的空病毒、miR-126mimic以及miR-126inhibitor的慢病毒。测定各组大鼠心肌梗死范围,TUNEL测定心肌细胞凋亡。结果:与对照组相比,心肌细胞H9C2SI/R后miR-126表达(0.32±0.02)明显降低(P=0.000)、细胞凋亡率(3.8±0.6)明显上升(P=0.000);与SI/R相比,采用miR-126mimic上调miR-126表达后细胞凋亡率(5.3±0.8)明显增加(P=0.007),caspase-3表达(0.450±0.003)明显降低(P=0.000),而采用miR-126inhibitor下调miR-126表达后细胞凋亡率(2.9±0.5)明显降低(P=0.000),caspase-3表达(1.80±0.03)明显升高(P=0.002)。与假手术组相比,I/R组和I/R+GFP组心肌梗死范围[(59.4±2.5),(60.2±2.8)]明显增加(P=0.000,P=0.000),而与I/R组相比,I/R+miR-126mimic组心肌梗死范围(69.0±2.4)明显增加(P=0.013),而I/R+miR-126inhibitor组心肌梗死范围(39.5±2.5)明显降低(P=0.000);TUNEL检测结果显示,与假手术组相比,I/R组和I/R+GFP组心肌细胞凋亡率[(33.4±1.7),(33.7±1.8)]明显升高(P=0.000,P=0.000);而与I/R组相比,I/R+miR-126mimic组心肌细胞凋亡率(42.2±2.3)明显增加(P=0.003),而I/R+miR-126inhibitor组心肌细胞凋亡率(25.5±1.5)明显降低(P=0.000)。结论:心肌缺血再灌注损伤会引起miR-126表达降低,而上调miR-126表达会促进心肌细胞凋亡,相反抑制miR-126表达可能有助于预防心肌缺血再灌注引起的心肌细胞凋亡。
[Abstract]:Aim: to investigate the role and mechanism of micro RNA(miR)-126 in myocardial ischemia reperfusion injury in vivo and in vitro. Methods: in vitro, myocardial H9C2 was divided into 6 groups to establish myocardial cell model of simulated ischemia / reperfusion SI / R injury, and then transfect miR-126mimic and miR-126inhibitor.qRT-PCR to detect the expression of miR-126 in cardiomyocytes. Apoptosis was detected by flow cytometry and the protein expression of caspase-3 and its fragmentation fragments were detected by Western blot. In vivo, healthy female Wistar rats were divided into 5 groups to establish ischemia / reperfusion injury model of ischemia / reperfusion injury, and transfect empty virus miR-126 mimic containing GFP and lentivirus containing miR-126inhibitor respectively. Myocardial apoptosis was determined by Tunel. Results: compared with the control group, the expression of miR-126 in cardiomyocytes after H9C2SI/R was 0.32 卤0.02) and the apoptosis rate was 3.8 卤0.6). When miR-126mimic was used to up-regulate the expression of miR-126, the apoptotic rate was 5.3 卤0.8) and the expression of caspase-3 was increased significantly (0.450 卤0.003). However, when miR-126inhibitor was used to down-regulate the expression of miR-126, the rate of cell apoptosis was 2.9 卤0.5), and the expression of caspase-3 was significantly decreased by 1.80 卤0.03) and the expression of caspase-3 was increased by 1.80 卤0.03. Compared with the sham operation group, the myocardial infarction size of I / R / R group and I / R GFP group [59.4 卤2.5 GFP 60.2 卤2.8] increased significantly, but the myocardial infarction size of I / R miR-126mimic group increased 69.0 卤2.4 compared with that of the I / R group, while the myocardial infarction size of I / R miR-126inhibitor group decreased 39.5 卤2.5), the results of Tunel showed that the myocardial infarction size of the I / R miR-126inhibitor group was 39.5 卤2.5, and that of the I / R miR-126inhibitor group was significantly lower than that of the I / R group, and that of the I / R miR-126mimic group was significantly higher than that of the I / R group. Compared with the sham operation group, the cardiomyocyte apoptosis rate in the I / R miR-126mimic group and the I / R GFP group [33.4 卤1.7 卤1.8] was significantly increased, while that in the I / R miR-126mimic group was significantly higher than that in the I / R miR-126mimic group (42.2 卤2.3), while in the IR miR-126inhibitor group, the myocardial cell apoptosis rate was 25.5 卤1.5) significantly lower than that in the I / R miR-126mimic group (25.5 卤1.5). Conclusion: myocardial ischemia-reperfusion injury can decrease the expression of miR-126, but up-regulating the expression of miR-126 can promote the apoptosis of myocardial cells. On the contrary, inhibiting the expression of miR-126 may help to prevent myocardial ischemia-reperfusion induced cardiomyocyte apoptosis.
【作者单位】：湖北医药学院附属东风医院心血管内科;
【基金】：湖北省自然科学基金重点资助项目(编号:2060402-791)
【分类号】：R54

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