miR-328-3p对氧化型低密度脂蛋白诱导的冠状动脉内皮细胞损伤的保护作用及机制研究  

Protective Effect and Mechanism of miR-328-3p on Coronary Artery Endothelial Cell Injury Induced by Oxidized Low-density Lipoprotein

作　　者：侯永兰[1] 李霞[1] 王建美 刘振[1] 韩明磊[1] 刘烝昊 金卫东[1] HOU Yonglan;LI Xia;WANG Jianmei;LIU Zhen;HAN Minglei;LIU Zhenghao;JIN Weidong(Department of Cardiovascular Medicine,Xinxiang Central Hospital,Xinxiang 453000,China)

机构地区：[1]新乡市中心医院心血管内科,新乡453000

出　　处：《四川大学学报（医学版）》2024年第5期1210-1216,共7页Journal of Sichuan University(Medical Sciences)

基　　金：河南省医学科技攻关计划联合共建项目(No.LHGJ20220987)资助。

摘　　要：目的探讨miR-328-3p对氧化型低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导的冠状动脉内皮细胞损伤的保护作用及可能相关的作用机制。方法用ox-LDL诱导人冠状动脉内皮细胞(human coronary artery endothelial cells,HCAECs),将细胞分为对照(control)组(正常培养细胞)、ox-LDL组(ox-LDL处理)、ox-LDL+miR-NC组(转染miR-NC,用ox-LDL处理)、ox-LDL+miR-328-3p组(转染miR-328-3p,用ox-LDL处理)、ox-LDL+miR-328-3p+pcDNA组(共转染miR-328-3p和pcDNA,用ox-LDL处理)、ox-LDL+miR-328-3p+胰岛素样生长因子2(insulin-like growth factor 2,IGF2)组(共转染miR-328-3p和IGF2,用ox-LDL处理)。RT-qPCR检测miR-328-3p表达水平;MTT以及流式细胞术检测细胞增殖和凋亡;Western blot法检测cleaved cas-3、IGF2、Bax、Bcl-2蛋白含量;ELISA法检测肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)、白细胞介素(interleukin,IL)-6、IL-1β含量;应用双荧光素酶报告实验验证分子间靶向关系。结果与control组相比,ox-LDL组中miR-328-3p表达水平、细胞活性降低(P<0.05),凋亡率,cleaved cas-3、IGF2蛋白表达,TNF-α、IL-6、IL-1β水平增加(P<0.05)。与ox-LDL+miR-NC组相比,ox-LDL+miR-328-3p组miR-328-3p表达水平、细胞活性增加(P<0.05),凋亡率,cleaved cas-3、IGF2蛋白表达,TNF-α、IL-6、IL-1β水平降低(P<0.05)。IGF2是miR-328-3p的功能靶标。与共转染oxLDL+miR-328-3p+pcDNA组比较,共转染ox-LDL+miR-328-3p+IGF2组IGF2蛋白水平升高(P<0.05),细胞活性降低(P<0.05),而凋亡率、cleaved cas-3蛋白水平以及TNF-α、IL-6、IL-1β含量升高(P<0.05)。结论miR-328-3p通过靶向负调控IGF2抑制ox-LDL诱导的冠状动脉内皮细胞凋亡和炎性损伤。Objective To investigate the protective effect of miR-328-3p on oxidized low-density lipoprotein(ox-LDL)-induced coronary artery endothelial cell injury and the potentially relevant mechanisms.Methods Human coronary artery endothelial cells(HCAECs)were induced with ox-LDL,and the cells were divided into a control group consisting of normal cells,an ox-LDL group receiving ox-LDL treatment,an ox-LDL+miR-NC group transfected with miR-NC and treated with ox-LDL,an ox-LDL+miR-328-3p group transfected with miR-328-3p and treated with ox-LDL,and ox-LDL+miR-328-3p+pcDNA group co-transfected miR-328-3p and pcDNA and treated with ox-LDL,and an ox-LDL+miR-328-3p+insulin-like growth factor 2(IGF2)group co-transfected miR-328-3p and IGF2 and treated with ox-LDL.The expression level of miR-328-3p was determined with RT-qPCR.Cell proliferation was determined by MTT.Cell apoptosis was measured by flow cytometry.Western blot was conducted to examine the protein expression levels of cleaved cas-3 and IGF2.ELISA was performed to determine the levels of tumor necrosis factorα(TNF-α),interleukin(IL)-6,and IL-1β.Dual luciferase reporter experiment was performed to verify the targeting relationship between miR-328-3p and IGF2.Results Compared with those of the control group,miR-328-3p expression level and cell activity were significantly reduced in the ox-LDL group(P<0.05),while the apoptotic rate,the protein expression levels of cleaved cas-3,IGF2,Bax,and Bcl-2,and the levels of TNF-α,IL-6,and IL-1βwere significantly increased(P<0.05).Compared with those of the ox-LDL+miR-NC group,miR-328-3p expression level and cell activity significantly increased in the ox-LDL+miR-328-3p group(P<0.05),while the apoptosis rate,the protein expression levels of cleaved cas-3 and IGF2,and the levels of TNF-α,IL-6,and IL-1βwere significantly reduced.IGF2 was a functional target of miR-328-3p.Compared with those of the ox-LDL+miR-328-3p+pcDNA co-transfection group,the IGF2 protein level was significantly increased(P<0.05)and cell activity was sign

关 键 词：miR-328-3p 胰岛素样生长因子2 氧化型低密度脂蛋白 冠状动脉内皮细胞 

分 类 号：R54[医药卫生—心血管疾病]