miR-19b对冠心病大鼠血管内皮细胞损伤的影响及PTEN/PI3K/AKT信号通路的作用  

Effect of miR-19b on vascular endothelial cell injury and the role of PTEN/PI3K/AKT signaling pathway in rats with coronary heart disease

作　　者：何娟 蒋肖潇 何秀波[1] HE Juan;JIANG Xiaoxiao;HE Xiubo(Department of Ultrasound Medicine,the Fourth Hospital of Changsha,Changsha,Hunan 410006,China)

机构地区：[1]湖南省长沙市第四医院超声医学科,湖南长沙410006

出　　处：《检验医学与临床》2024年第18期2657-2663,2669,共8页Laboratory Medicine and Clinic

摘　　要：目的探讨微小RNA(miR)-19b对冠心病大鼠血管内皮细胞损伤的影响及磷酸酶及张力蛋白同源物/磷脂酰肌醇3-激酶/蛋白激酶B(PTEN/PI3K/AKT)信号通路的作用。方法选取72只大鼠随机分为正常组、模型组、模拟对照物(NC)组、miR-19b组、miR-19b+质粒互补DNA(pcDNA)3.1组、miR-19b+PTEN组。采用高脂饲料+垂体后叶素建立冠心病模型,尾静脉注射miR-19b mimic、pcDNA3.1-PTEN慢病毒液构建miR-19b、PTEN过表达大鼠。采用荧光定量聚合酶链反应检测冠状动脉组织miR-19b和PTEN信使RNA(mRNA)水平,测定大鼠心功能,观察大鼠冠状动脉病理变化,检测心肌组织细胞凋亡情况、血管内皮功能、冠状动脉中PTEN/PI3K/AKT信号通路、含半胱氨酸的天冬氨酸蛋白水解酶(cleaved Caspase)-3、cleaved Caspase-9水平。分离培养冠心病大鼠冠状动脉内皮细胞,根据转染方式分为野生型质粒(WT)+NC组、WT+miR-19b mimic组、突变型质粒(MUT)+NC组、MUT+miR-19b mimic组,采用双荧光素酶报告基因实验验证miR-19b与PTEN的靶向关系。将冠心病大鼠冠状动脉内皮细胞分为正常细胞组、NC细胞组和miR-19b细胞组,检测PTEN mRNA和PTEN蛋白水平。结果miR-19b组左心室射血分数(LVEF)、miR-19b、血清血管内皮生长因子(VEGF)、一氧化氮(NO)、磷酸化磷脂酰肌醇-3激酶(p-PI3K)/PI3K蛋白、磷酸化蛋白激酶B(p-AKT)/AKT蛋白水平高于模型组,左心室收缩末期容积(LVESV)、舒张末期容积(LVEDV)、细胞凋亡率、cleaved Caspase-3和cleaved Caspase-9、PTEN mRNA、内皮素-1(ET-1)、PTEN蛋白水平低于模型组,差异均有统计学意义(P<0.05)。miR-19b+PTEN组LVESV、LVEDV、细胞凋亡率、PTEN mRNA、cleaved Caspase-3、cleaved Caspase-9、ET-1、VEGF、PTEN蛋白水平高于miR-19b组,LVEF、NO、p-PI3K/PI3K蛋白和p-AKT/AKT蛋白水平低于miR-19b组,差异均有统计学意义(P<0.05)。模型组LVEF、miR-19b、VEGF、NO、p-PI3K/PI3K蛋白、p-AKT/AKT蛋白水平低于正常组,LVESV�Objective To investigate the effects of microRNA(miR)-19b on vascular endothelial cell injury in rats with coronary heart disease and the role of phosphatase and tensin homology/phosphatidylinositol 3-kinase/protein kinase B(PTEN/PI3K/AKT)signaling pathway.Methods A total of 72 rats were randomly divided into normal group,model group,simulated control(NC)group,miR-19b group,miR-19b+plasmid complementory DNA(pcDNA)3.1 group and miR-19b+PTEN group.The coronary heart disease model was established using high-fat diet and pituitin.miR-19b and PTEN overexpression rats were constructed by tail vein injection of miR-19b mimic and pcDNA3.1-PTEN lentivirus solution.Fluorescence quantitative polymerase chain reaction was used to detect the levels of miR-19b and PTEN messenger RNA(mRNA)in coronary tissue,determine the cardiac function of rats and observe the pathological changes of rat coronary arteries.Apoptosis of myocardial tissue cells,vascular endothelial function,PTEN/PI3K/AKT signaling pathway and cleaved cysteinyl aspartate specific proteinase-3(cleaved Caspase-3)and cleaved Caspase-9 levels after cleavage in coronary arteries were detected.The coronary endothelial cells of coronary heart disease rats were isolated and cultured.They were divided into wild-type plasmid(WT)+NC group,WT+miR-19b mimic group,mutant plasmid(MUT)+NC group and MUT+miR-19b mimic group according to transfection mode.Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-19b and PTEN.The coronary endothelial cells of coronary heart disease rats were divided into normal cell group,NC cell group and miR-19b cell group,and the levels of PTEN mRNA and PTEN protein were detected.Results The levels of left ventricular ejection fraction(LVEF),miR-19b,serum vascular endothelial growth factor(VEGF),nitric oxide(NO),phosphorylated phosphatidylinositol 3-kinase(p-PI3K)/PI3K protein and phosphorylated protein kinase B(p-AKT)/AKT protein in miR-19b group were higher than those in model group,and left ventricular end-sy

关 键 词：冠心病 微小RNA-19b 磷酸酶及张力蛋白同源物 磷脂酰肌醇3-激酶 蛋白激酶B 内皮细胞损伤 

分 类 号：R541.4[医药卫生—心血管疾病] R363.21[医药卫生—内科学]