摘要
目的:探讨miR-140-3p靶向调节PI3K/Akt通路对缺氧复氧心肌细胞再灌注损伤的影响。方法:对大鼠来源的H9C2心肌细胞进行缺氧/复氧以诱导细胞损伤,使用miR-140-3p mimics及其对照(NC)转染H9C2心肌细胞,再使用10μmol/L LY294002处理过表达miR-140-3p的H9C2细胞。将心肌细胞分为6组,分别为对照组(CON组)、H/R组(Model组)、Model+NC组、Model+miR-140-3p mimics组、Model+miR-140-3p mimics+LY294002组、Model+LY294002组。采用荧光定量PCR(RT-PCR)法检测各组miR-140-3p相对表达量,CCK-8法检测各组心肌细胞活力,流式细胞仪检测各组心肌细胞凋亡率,Western blot检测各组磷酸化磷脂酰肌醇3激酶(p-PI3K)、磷酸化蛋白激酶B(p-Akt)、Bax、Bcl-2、Cleaved-caspases-3表达量。结果:与CON组比较,Model组miR-140-3p表达量、细胞活力以及Bcl-2、p-PI3K、p-Akt蛋白表达明显降低(P<0.05),Bax、Cleaved-caspases-3蛋白表达和凋亡率明显提高(P<0.05),与Model组比较,miR-140-3p过表达可上调miR-140-3p表达量、细胞活力以及Bcl-2、p-PI3K、p-Akt蛋白,下调Bax、Cleaved-caspases-3蛋白表达和凋亡率,LY294002可抑制miR-140-3p表达,下调细胞活力以及Bcl-2、p-PI3K、p-Akt蛋白,上调Bax、Cleaved-caspases-3蛋白表达和凋亡率。结论:MiR-140-3p可通过调节PI3K/Akt通路减轻缺氧复氧心肌细胞自噬和凋亡,进而减轻心肌损伤。
Objective:To investigate the effect of miR-140-3p on hypoxia/reoxygenation-induced myocardial cell reperfusion injury by targeting PI3K/Akt pathway.Methods:H9C2 cardiomyocytes derived from rats were subjected to hypoxia/reoxygenation to induce cell injury.MiR-140-3p mimics and their control(NC)were transfected into H9C2 cardiomyocytes,and H9C2 cells overexpressing miR-140-3p were treated with 10μmol/L LY294002.The cardiomyocytes were divided into 6 groups,named control group(CON group),H/R group(Model group),Model+NC group,Model+miR-140-3p mimics group,Model+miR-140-3p mimics+LY294002 group,Model+LY294002 group.The relative expression of miR-140-3 p in each group was detected by fluorescence quantitative PCR(RT-PCR).The viability of cardiomyocytes in each group was detected by CCK-8 method.The apoptosis rate of cardiomyocytes in each group was detected by flow cytometry.The expression levels of phosphorylated phosphatidylinositol 3 kinase(p-PI3K),phosphorylated protein kinase B(p-Akt),Bax,Bcl-2,and cleared caspases-3 in each group were detected by Western blot.Results:Compared with the CON group,the expression level and cell viability of miR-140-3p,as well as the expression of Bcl-2,p-PI3K,and p-Akt proteins,were significantly reduced in the Model group(P<0.05),while the expression and apoptosis rate of Bax and Cleared caspases-3 proteins were significantly increased(P<0.05).Compared with the Model group,overexpression of miR-140-3p upregulated the expression level and cell viability of miR-140-3p,as well as Bcl-2,p-PI3K,and p-Akt proteins,while downregulated the expression and apoptosis rate of Bax and Cleared caspases-3 proteins,LY294002 could inhibit the expression of miR-140-3p,downregulate cell viability and Bcl-2,p-PI3K,p-Akt proteins,and upregulate the expression of Bax,Cleared caspases-3 proteins and apoptosis rate.Conclusion:MiR-140-3p can alleviate autophagy and apoptosis in hypoxia/reoxygenation cardiomyocytes by regulating the PI3K/Akt pathway,thereby alleviating myocardial injury.
作者
郑伟
蔺雪峰
韩轩茂
李阳
ZHENG Wei;LIN Xuefeng;HAN Xuanmao;LI Yang(Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014040,China;The First Department of Cardiology,the First Affiliated Hospital of Baotou Medical College of Inner Mongolia University of Science and Technology)
出处
《包头医学院学报》
CAS
2024年第8期26-31,共6页
Journal of Baotou Medical College
基金
内蒙古自治区自然科学基金项目(2023MS08068)。
作者简介
通讯作者:蔺雪峰。
