摘要
目的探讨miR-7在心肌细胞缺氧复氧模型中的表达及对心肌细胞凋亡的影响。方法构建心肌细胞缺氧复氧模型,RT-PCR检测心肌细胞中miR-7的表达水平。心肌细胞转染miR-7模拟物和抑制物,RT-PCR检测转染效果。流式细胞术检测miR-7模拟物和抑制物对缺氧复氧心肌细胞凋亡的影响,试剂盒检测细胞中乳酸盐脱氢酶(LDH)水平,Western blot检测细胞中活化的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved caspase-3)、磷酸化蛋白激酶B(p-Akt)蛋白水平。结果缺氧复氧心肌细胞模型中miR-7的表达水平明显高于正常培养的心肌细胞(P<0.01)。miR-7模拟物能够提高心肌细胞中miR-7的表达水平,而miR-7抑制物能够抑制心肌细胞中miR-7的表达。miR-7模拟物能够降低缺氧复氧心肌细胞的凋亡率和细胞中cleaved caspase-3表达,抑制心肌细胞分泌LDH,促进细胞中p-Akt的表达。miR-7抑制物能够促进缺氧复氧心肌细胞凋亡和细胞中cleaved caspase-3表达,促进心肌细胞分泌LDH,抑制细胞中p-Akt的表达。结论 miR-7在心肌细胞缺氧复氧模型中表达升高。miR-7能够降低缺氧复氧心肌细胞凋亡,干扰miR-7表达能够促进缺氧复氧心肌细胞凋亡,作用机制可能与Akt信号通路有关。
Objective To explore the expression of miR-7 in myocardial hypoxia reoxygenation model and its effect on the ap- optosis of myocardial cells. Methods Myocardial hypoxia reoxygenation model was established, and the expression level of miR-7 in myocardial cells was detected by reverse transcription polymerase chain reaction （ RT-PCR}. Myocardial cells were transfeeted with miR-7 mimics and inhibitors, and then RT-PCR was used to detect the transfection effect. Flow cytometry was employed to detect the effects of miR-7 mimics and inhibitors on myocardial apoptosis induced by hypoxia reoxygenation. Lactate dehydrogenase （LDH） kit was used to detect the LDH level, and the protein expression levels of cleaved caspase-3 and p-Akt in cells were detected by Western blot. Results The expression level of miR-7 in hypoxia reoxygenation myocardial cells was sig- nificantly higher than that of normal cultured cardiomyocytes （ P〈0.01 ）. miR-7 mimics could enhance the expression level of miR -7 in myocardial cells, while miR-7 inhibitors could inhibit the expression of miR-7 in myocardial ceils, miR-7 mimics could re- duce the apoptosis rate of hypoxia reoxygenation myocardial cells and the expression of cleaved caspase-3 in the cells, inhibit the secretion of LDH in cardiac myocytes and promote the expression of p-Akt in the cells, miR-7 inhibitors could promote the apopto- sis rate of hypoxia reoxygenation myocardial cells, increase the expression of cleaved easpase-3 in the cells, promote the secretion of LDH in cardiac myocytes and inhibit the expression of p-Akt in the cells. Conclusions The expression of miR-7 in myocar- dial hypoxia reoxygenation model is elevated, miR-7 can reduce the apoptosis of hypoxia reoxygenation myocardial cells, interfer- ence of miR-7 expression can promote the apoptosis of hypoxia reoxygenation myocardial cells, and the mechanism may be related to Akt signaling pathway.
出处
《实用预防医学》
CAS
2018年第2期195-198,共4页
Practical Preventive Medicine
基金
上海市浦东新区计划项目(No:PWRd2016-19)
关键词
心肌细胞
缺氧复氧
凋亡
miR-7
myocardial cell
hypoxia reoxygenation
apoptosis
miR-7
