微小RNA-423-5p靶向乙醛脱氢酶2减轻脂多糖诱导的人脐静脉血管内皮细胞损伤  

MicroRNA-423-5p reducing the injury of human umbilical vascular endothelial cells induced by lipopolysaccharide through targeting aldehyde dehydrogenase 2

作　　者：刘文芳 LIU Wen-fang(Teaching and Research Office of Biochemistry and Hygiene,Linyi Health Schoob,Shandong Province,Shandong Linyi276000,China)

机构地区：[1]山东省临沂卫生学校生物生化卫生学教研室,山东临沂276000

出　　处：《解剖学报》2020年第6期868-876,共9页Acta Anatomica Sinica

摘　　要：目的探讨微小RNA-423-5p(miR-423-5p)对脂多糖(LPS)诱导血管内皮细胞损伤的保护及作用机制。方法用1 mg/L LPS诱导人脐静脉血管内皮细胞(HUVECs)24 h,Real-time PCR和Western blotting检测细胞中miR-423-5p和乙醛脱氢酶2(ALDH2)的表达。通过转染anti-miR-423-5p和pc DNA-ALDH2下调miR-423-5p和上调ALDH2表达,流式细胞术检测细胞凋亡率,Western blotting检测凋亡相关蛋白Bcl-2和Bax的表达,并用ELISA试剂盒检测LPS诱导后细胞上清液中白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)的含量;双荧光素酶报告系统验证miR-423-5p与ALDH2的调控关系。结果与对照相比,LPS可诱导HUVECs凋亡和损伤,使HUVECs中miR-423-5p、Bax表达量及IL-6和TNF-α分泌量均显著升高(P<0.05),ALDH2的mRNA和蛋白表达量及Bcl-2量显著降低(P<0.05);下调miR-423-5p表达和过表达ALDH2均可减轻LPS诱导的HUVECs损伤并抑制细胞凋亡;miR-423-5p靶向负调控ALDH2的表达;抑制ALDH2表达逆转了下调miR-423-5p表达对LPS诱导的HUVECs损伤的作用。结论下调miR-423-5p表达可靶向ALDH2减轻LPS对HUVECs的损伤并抑制细胞凋亡。Objective To investigate the protective effect of microRNA-423-5 p( miR-423-5 p) on injury of human umbilical vein endothelial cells( HUVECs) induced by lipopolysaccharide( LPS) and its potential mechanism. Methods The HUVECs were induced by 1 mg/L LPS for 24 hours,then expression levels of miR-423-5 p and aldehyde dehydrogenasel2( ALDH2) in HUVECs induced by LPS were determined by Real-time PCR and Western blotting. The miR-423-5 p was down-regulated and ALDH2 was up-regulated by transfected with anti-miR-423-5 p and pc DNA-ALDH2. The apoptotic rate of HUVECs was detected by flow cytometry,and the expression levels of Bcl-2 and Bax were measured by Western blotting.The levels of interleukin-6( IL-6) and tumor necrosis factor α( TNF-α) in supernatant of HUVECs induced by LPS were determined by the kits. The relationship between miR-423-5 p and ALDH2 was validated by dual-luciferase reporter assay system. Results Compared with the control group,LPS induced apoptosis and injury of HUVECs. After LPS induction,the levels of miR-423-5 p,Bax,IL-6 and TNF-α in HUVECs increased significantly( P < 0. 05),and the levels of Bcl-2,ALDH2 mRNA and ALDH2 protein decreased remarkably( P < 0. 05). Either down-regulation of miR-423-5 p or overexpression of ALDH2 alleviated the injury of HUVECs induced by LPS and inhibited apoptosis. MiR-423-5 p targeted and negatively regulated the expression of ALDH2. Inhibition of ALDH2 reversed the effects of miR-423-5 p down-regulation on the injury of HUVECs induced by LPS. Conclusion MiR-423-5 p alleviates the injury of HUVECs induced by LPS and inhibits apoptosis by targeting ALDH2.

关 键 词：脐静脉血管内皮细胞 脂多糖 微小RNA-423-5p 乙醛脱氢酶2 实时定量聚合酶反应 流式细胞术 人 

分 类 号：R543.6[医药卫生—心血管疾病]