miR-323-3p通过靶向TGF-β2/JNK通路影响冠心病大鼠心肌细胞的凋亡  

Effect of miR-323-3p on cardiomyocyte apoptosis in rats with coronary heart disease via targeting TGF-β2/JNK pathway

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;WANG Zongshe;SHU Ruichao;DUAN Zhaoyang;MA En(Baoji Central Hospial,Baoji 721000,China;The Second Affliated Hospital of Xian Jiaotong University,Xian 710004,China)

机构地区：[1]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：《现代医学》2022年第7期840-846,共7页Modern Medical Journal

基　　金：宝鸡市卫生健康委员会资助项目(2020-001)。

摘　　要：目的:探讨微小RNA-323-3p(miR-323-3p)对冠心病大鼠心肌细胞凋亡的影响及其可能的作用机制。方法:将SD大鼠分为Control组、Model组、NC agomir组、miR-323-3p agomir组、miR-323-3p agomir+pcDNA组、miR-323-3p agomir+pcDNA-TGF-β2组,每组12只。除Control组外,其它组大鼠均采用高脂饲料喂养以构建冠心病大鼠模型。建模成功后,通过尾静脉注射对应的agomir或pcDNA进行处理,Control组、Model组尾静脉注射等量的生理盐水。2周后,测量大鼠心脏功能指标平均动脉压(MAP)、左室收缩压(LVSP)、心率(HR)的变化;HE染色检测大鼠心肌组织病理变化;TUNEL染色检测各组大鼠心肌细胞凋亡;qRT-PCR检测大鼠心肌组织中miR-323-3p表达;Western blotting检测大鼠心肌组织中Bax、Bcl-2、转化生长因子β2(TGF-β2)及磷酸化c-Jun氨基末端激酶(p-JNK)蛋白表达;双荧光素酶报告基因实验验证miR-323-3p与TGF-β2的关系。结果:与Control组比较,Model组大鼠心肌细胞肿胀,有大量炎症细胞浸润,心肌纤维断裂,MAP、LVSP、HR、miR-323-3p、Bcl-2蛋白表达水平显著降低,细胞凋亡率、Bax、TGF-β2、p-JNK蛋白表达水平显著升高(P<0.05);与Model组、NC agomir组比较,miR-323-3p agomir组大鼠心肌组织病理改变显著减轻,MAP、LVSP、HR、miR-323-3p、Bcl-2蛋白表达水平显著升高,细胞凋亡率、Bax、TGF-β2、p-JNK蛋白表达水平显著降低(P<0.05);与miR-323-3p agomir组、miR-323-3p agomir+pcDNA组比较,miR-323-3p agomir+pcDNA-TGF-β2组大鼠心肌组织病理改变显著加重,MAP、LVSP、HR、Bcl-2蛋白表达水平显著降低,细胞凋亡率、Bax、TGF-β2、p-JNK蛋白表达水平显著升高(P<0.05),而miR-323-3p水平变化差异无统计学意义(P>0.05);miR-323-3p靶向调控TGF-β2的表达。结论:过表达miR-323-3p可能通过抑制TGF-β2/JNK通路抑制冠心病大鼠细胞凋亡。Objective: To investigate the effect of miRNA-323-3 p on cardiomyocyte apoptosis in rats with coronary heart disease and its possible mechanism. Methods: The SD rats were divided into Control group, Model group, NC agomir group, miR-323-3 p agomir group, miR-323-3 p agomir+pcDNA group, and miR-323-3 p agomir+pcDNA-TGF-β2 group, 12 rats in each group. Except for the control group, rats in the other groups were fed with high-fat diet to construct a coronary heart disease rat model. After the modeling was successfully estabilshed, the corresponding agomir or pcDNA was injected into the tail vein for treatment, the control group and the model group were injected with the same amount of normal saline through the tail vein. Two weeks later, the changes in mean arterial pressure(MAP), left ventricular systolic pressure(LVSP), and heart rate(HR) of the rats’ heart function indexes were measured;HE staining was used to detect the pathological changes of rat myocardial tissue;TUNEL staining was used to detect myocardial cell apoptosis in rats of each group;qRT-PCR was used to detect the miR-323-3 p expression in rat myocardial tissue;Western blotting was used to detect the Bax, Bcl-2, transforming growth factor β2(TGF-β2) and phosphorylated c-Jun N-terminal kinase(p-JNK) protein expression in rat myocardial tissue;and dual luciferase reporter gene experiment was used to verify the relationship between miR-323-3 p and TGF-β2. Results: Compared with the control group, the myocardial cells were swollen, with a large number of inflammatory cells infiltrating and myocardial fibers breaking, the MAP, LVSP, HR, and the miR-323-3 p and Bcl-2 protein expression levels reduced significantly, and apoptosis rate, Bax, TGF-β2 and p-JNK protein expression levels increased significantly in the model group(P<0.05);compared with the model group and NC agomir group, the pathological changes of myocardial tissue were significantly reduced, the MAP, LVSP, HR, the miR-323-3 p and Bcl-2 protein expression levels increased significantly,

关 键 词：miR-323-3p 冠心病 细胞凋亡 转化生长因子β2/c-Jun氨基末端激酶通路 大鼠 

分 类 号：R-33[医药卫生] R541.4