%0 Journal Article
%T miR-499对缺氧/复氧诱导的心肌细胞凋亡的影响<br>Effect of miR-499 on apoptosis of primary cardiomyocytes induced by anoxia-reoxygenation
%A 岳
%A 莹
%A 吕风华
%A 陈玉磊
%A 王
%A 卓
%A 司澳洋
%J 郑州大学学报（医学版）
%D 2018
%R 10.13705/j.issn.1671-6825.2017.11.149
%X 目的:探讨miR-499对缺氧/复氧诱导的原代心肌细胞凋亡的影响,探讨其与磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)通路的关系。方法:通过对原代培养新生大鼠心肌细胞行缺氧3 h/复氧4 h,模拟大鼠心肌缺血再灌注损伤; 将细胞分为4组:正常对照组、缺氧/复氧组(A/R组)、缺氧/复氧+miR-499 激动剂agomir预处理组(Agomir组)、缺氧/复氧+miR-499激动剂 agomir+PI3K特异性阻断剂LY294002组(Agomir+LY294002组)。采用MTT方法检测各组心肌细胞活力,分别采用荧光定量PCR方法和Western blot法检测各组心肌细胞中miR-499、PI3K mRNA及Caspase-3、p-PI3K和p-Akt蛋白表达水平的变化。结果:与正常对照组相比,A/R组心肌细胞活力下降,miR-499及PI3K mRNA表达量减少,p-PI3K及p-Akt蛋白表达量减少,Caspase-3蛋白表达量增加(P<0.05); 但经 Agomir处理后,心肌细胞活力增加,miR-499及PI3K mRNA表达量增加; p-PI3K及p-Akt蛋白表达量增加; 与A/R组相比,Caspase-3蛋白表达量减少(P<0.05); 而agomir的作用可被LY294002即PI3K阻断剂抑制。结论:miR-499可以减少缺氧/复氧所诱导的心肌细胞的凋亡,这可能与其活化PI3K/Akt信号通路有关。<br>Aim: To investigate the effect of miR-499 on apoptosis of primary cardiomyocytes induced by anoxia-reoxygenation(A/R), and its potential relationship with PI3K/Akt pathway.Methods: The cultured primary cardiomyocytes from neonatal rats were divided into 4 groups: control group, anoxia-reoxygenation group(A/R group), anoxia-reoxygenation+miR-499 agomir group(Agomir group), anoxia-reoxygenation+miR-499 agomir+LY294002 group(Agomir+LY294002 group). The procedure of A/R was performed in cultured primary cardiomyocytes by 3 h anoxia and then 4 h reoxygenation. The viability of the cells was detected by MTT assay. The expressions of miR-499 and PI3K mRNA were detected by RT-PCR. The protein expression levels of Caspase-3, phosphorylated Akt(p-Akt), and phosphorylated PI3K(p-PI3K)were measured by Western blot assay.Results: Compared with control group, the cell viability, expressions of miR-499 and PI3K mRNA, and the protein expression levels of p-Akt and p-PI3K were significantly decreased, and the protein expression level of Caspase-3 was increased in A/R group(P<0.05). Compared with A/R group, the protein expression level of Caspase-3 was markedly decreased, while the cell viability, the expression of miR-499 and PI3K mRNA as well as the protein expression levels of p-Akt and p-PI3K were significantly increased in Agomir group(P<0.05). The effect of miR-499 agomir was inhibited by LY294002.Conclusion: miR-499 could protect the primary cardiomyocytes from A/R-induced apoptosis, which may be involved in the activation of PI3K/Akt signaling pathway
%K miR-499
%K 缺氧/复氧
%K 细胞凋亡
%K 磷脂酰肌醇3-激酶/蛋白激酶B通路&lt
%K br&gt
%K miR-499
%K anoxia/reoxygenation
%K apoptosis
%K PI3K/Akt pathway
%U http://jms.zzu.edu.cn/oa/darticle.aspx?type=view&id=201804027