MiR-362-3p靶向调控ORM1对缺氧复氧心肌细胞增殖及凋亡影响  

Influence of miR-362-3p on proliferation and apoptosis of hypoxia/reoxygenation cardiomyocytes through targeting and controlling α1-acid glycoprotein 1

作　　者：张飞飞[1] 王文静[1] 谢悦陶[1] 李鑫柠[1] 史媚菁 党懿[1] Zhang Feifei;Wang Wenjing;Xie Yuetao;Li Xinning;Shi Meijing;Dang Yi(Department of Cardiovascular Medicine,People's Hospital of Hebei Province,Shijiazhuang 050051,China;不详)

机构地区：[1]河北省人民医院心血管内科,石家庄050051 [2]河北医科大学,石家庄050017

出　　处：《中国循证心血管医学杂志》2022年第4期476-479,483,共5页Chinese Journal of Evidence-Based Cardiovascular Medicine

摘　　要：目的 探讨微小RNA362-3p(miR-362-3p)靶向α-1-酸性糖蛋白(ORM1)对缺氧复氧(H/R)心肌细胞增殖及凋亡影响。方法 构建H/R损伤的心肌细胞模型,RT-qPCR检测miR-362-3p表达水平的变化,分别转染miR-362-3p mimic及miR-362-3p inhibitor,CCK-8法检测细胞增殖,流式细胞术测定细胞凋亡,通过TargetScan和双荧光素实验验证miR-362-3p和ORM1的靶向关系;Western blot检测miR-362-3p mimic及miR-362-3p inhibitor对ORM1、Caspase-9蛋白表达影响。结果 MiR-362-3p在H/R组心肌细胞中低表达(P<0.01);H/R组细胞增殖显著降低,细胞凋亡率明显增加(P<0.05),上调miR-362-3p可显著改善H/R后心肌细胞增殖及凋亡率(P<0.05),抑制miR-362-3p可进一步降低H/R后心肌细胞增殖,增加细胞凋亡率(P<0.05)。双萤光素酶报告实验提示miR-362-3p靶向作用于ORM1。H/R后心肌细胞ORM1、Caspase-9蛋白表达增加,上调miR-362-3p降低ORM1、Caspase-9蛋白表达,抑制miR-362-3p可增加Caspase-3蛋白表达。结论 MiR-362-3p可通过靶向ORM1改善缺氧复氧心肌细胞增殖及凋亡。Objective To discuss the influence of miR-362-3p on proliferation and apoptosis of hypoxia/reoxygenation(H/R) cardiomyocytes through targeting and controlling α1-acid glycoprotein 1(ORM1).Methods The model of H/R cardiomyocytes was established.The changes of miR-362-3p expression were detected by using RT-qPCR.H/R cardiomyocytes were transfected with miR-362-3p mimic and miR-362-3p inhibitor respectively,and proliferation of cardiomyocytes was detected by using CCK-8 and apoptosis of cardiomyocytes was detected by using flow cytometry.The targeting relationship between miR-362-3p and ORM1 was verified by using TargetScan and dual-luciferase reporter gene assay.The influences of miR-362-3p mimic and miR-362-3p inhibitor on expressions of ORM1 and Caspase-9 were detected by using Western blotting assay.Results The expression of miR-362-3p was lower in cardiomyocytes in H/R group(P<0.01).The cardiomyocyte proliferation decreased significantly and apoptosis rate increased significantly in H/R group(P<0.05).The up-regulation of miR-362-3p could significantly ameliorate proliferation and apoptosis rate of cardiomyocytes after H/R(P<0.05),and inhibition of miR-362-3p could reduce further cardiomyocyte proliferation and increase apoptosis rate of cardiomyocytes after H/R(P<0.05).The results of dual-luciferase reporter gene assay showed that miR-362-3p targeted and acted to ORM1.The expressions of ORM1 and Caspase-9 protein increased in cardiomyocytes after H/R.The up-regulation of miR-362-3p reduced expressions of ORM1 and Caspase-9 protein,and inhibition of miR-362-3p promoted expression of Caspase-3 protein.Conclusion MiR-362-3p can ameliorate proliferation and apoptosis of H/R cardiomyocytes through targeting ORM1.

关 键 词：微小RNA-362-3p α-1-酸性糖蛋白 缺氧 复氧 细胞凋亡 

分 类 号：R542.2[医药卫生—心血管疾病]