miR-383-3p靶向PTEN/PI3K/Akt/mTOR信号通路对冠心病大鼠内皮细胞凋亡的影响  

Effect of miR-383-3p Targeting PTEN/PI3K/Akt/mTOR Signaling Pathway on Endothelial Cell Apoptosis in Rats with Coronary Atherosclerotic Heart Disease

作　　者：蔡贵东 靳孟妮 王小军[1] 王宗社[1] 舒瑞朝 段朝阳[2] 马恩[1] CAI Guidong;JIN Mengni;WANG Xiaojun(Baoji Central Hospital,Shaanxi Baoji 721000,China)

机构地区：[1]陕西省宝鸡市中心医院,陕西宝鸡721000 [2]西安交通大学第二附属医院,陕西西安710004

出　　处：《河北医学》2022年第12期1943-1949,共7页Hebei Medicine

基　　金：陕西省宝鸡市卫生健康委员会资助项目,(编号:NO2020-001)。

摘　　要：目的:探究微小RNA-383-3p(miR-383-3p)靶向调节人第10号染色体缺失的磷酸酶/磷脂酰肌醇3-激酶/蛋白激酶B/雷帕霉素靶蛋白(PTEN/PI3K/AKT/mTOR)信号通路对冠心病(CHD)大鼠内皮细胞凋亡的影响。方法:将SD大鼠分为control组、CHD组、mimic NC组、miR-383-3p mimic组、miR-383-3p mimic+pcDNA3.1组、miR-383-3p mimic+pcDNA3.1-PTEN组,每组15只;除control组外其余组大鼠通过高脂饲料喂养及垂体后叶素注射构建CHD模型,造模前24h,将慢病毒液或质粒载体注射到相应组大鼠中;实时荧光定量PCR(qRT-PCR)检测冠状动脉miR-383-3p表达;全自动生化分析仪测定大鼠血脂水平;酶联免疫吸附(ELISA)法检测血清内皮素-1(ET-1)、血管紧张素Ⅱ(AngⅡ)、血管内皮生长因子(VEGF)、一氧化氮(NO)水平;HE染色及TUNEL法观察冠状动脉组织病理变化及内皮细胞凋亡情况;双荧光素酶报告基因实验验证miR-383-3p与PTEN靶向关系;Western blot检测冠状动脉组织Bcl-2、Bax及PTEN/PI3K/Akt/mTOR通路蛋白表达。结果:与control组相比,CHD组大鼠miR-383-3p、高密度脂蛋白胆固醇(HDL-C)、VEGF、NO、Bcl-2、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR水平显著减少,三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、总胆固醇(TC)、ET-1、AngⅡ水平、冠状动脉组织病理损伤程度、内皮细胞凋亡指数(AI)、Bax、PTEN表达显著升高(P<0.05);与CHD组相比,miR-383-3p mimic组miR-383-3p、HDL-C、VEGF、NO、Bcl-2、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR水平显著增加,TG、LDL-C、TC、ET-1、AngⅡ水平、冠状动脉组织病理损伤程度、内皮细胞AI、Bax、PTEN表达显著减少(P<0.05);过表达PTEN可逆转miR-383-3p高表达对CHD大鼠内皮细胞凋亡及功能损伤的改善作用;PTEN是miR-383-3p的靶向基因。结论:miR-383-3p可通过靶向抑制PTEN表达来激活PI3K/Akt/mTOR信号通路,与抑制CHD大鼠内皮细胞凋亡密切相关。Objective:To explore the effect of microRNA-383-3p(miR-383-3p)on endothelial cell apoptosis in rats with coronary atherosclerotic heart disease(CHD)by targeting the gene of phosphate and tension homology deleted on chromsome ten/Phosphoinositide 3-kinases/protein kinase B/mammalian target of rapamycin(PTEN/PI3K/AKT/mTOR)signaling pathway.Methods:The SD rats were divided into control group,CHD group,mimic NC group,miR-383-3p mimic group,miR-383-3p mimic+pcDNA3.1 group,and miR-383-3p mimic+pcDNA3.1-PTEN group,with 15 rats in each group;except for the control group,rats in the other groups were fed with high-fat diet and injected with pituitrin to construct a CHD model,and the lentivirus solution or plasmid vector was injected into the corresponding group of rats at 24 hours before modeling;quantitative Real-time(qRT-PCR)was used to detect the expression of miR-383-3p in coronary arteries;automatic biochemical analyzer was used to determine the level of blood lipid in rats;enzyme linked immunosorbent assay(ELISA)was used to detect the levels of serum endothelin-1(ET-1),angiotensinⅡ(AngⅡ),vascular endothelial growth factor(VEGF)and nitric oxide(NO);HE staining and TUNEL method were used to observe the pathological changes of coronary artery tissues and endothelial cell apoptosis;dual luciferase reporter gene experiment was used to verify the targeting relationship between miR-383-3p and PTEN;Western blot was used to detect the expression of Bcl-2,Bax and PTEN/PI3K/Akt/mTOR pathway protein in coronary artery tissue.Results:Compared with the control group,the miR-383-3p,high-density lipoprotein cholesterol(HDL-C),VEGF,NO,Bcl-2,p-PI3K/PI3K,p-Akt/Akt and p-mTOR/mTOR levels were significantly reduced in the CHD group,the triacylglycerol(TG),low-density lipoprotein cholesterol(LDL-C),total cholesterol(TC),ET-1,AngⅡlevels,coronary artery tissue pathological damage,endothelial apoptosis index(AI),Bax,PTEN expression were significantly increased(P<0.05);compared with the CHD group,the miR-383-3p,HDL-C,VEGF,NO,Bcl-2,p-PI

关 键 词：微小RNA-383-3p 人第10号染色体缺失的磷酸酶/磷脂酰肌醇3-激酶/蛋白激酶B/雷帕霉素靶蛋白 冠状动脉粥样硬化性心脏病 凋亡 

分 类 号：R541.4[医药卫生—心血管疾病]