茯苓酸调节miR-145-5p/KLF5轴对ox-LDL诱导的血管内皮细胞凋亡的影响  

Effect of Poric Acid on ox-LDL-induced Apoptosis of Vascular Endothelial Cells by Regulating miR-145-5p/KLF5 Axis

作　　者：吕静静 张溢寒 汪东东 LYU Jingjing;ZHANG Yihan;WANG Dongdong(Zhengzhou Hospital of Traditional Chinese Medicine,Zhengzhou 450006,Henan,China)

机构地区：[1]郑州市中医院,郑州450006

出　　处：《中西医结合心脑血管病杂志》2024年第20期3697-3703,共7页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：河南省中医药科学研究专项课题(No.2018ZY3008,2023ZY2151)。

摘　　要：目的:探讨茯苓酸调节miR-145-5p/Krüppel样转录因子5(KLF5)轴对氧化修饰的低密度脂蛋白(ox-LDL)诱导的血管内皮细胞凋亡的影响。方法:将人脐静脉血管内皮细胞(HUVEC)分为对照组、ox-LDL组、ox-LDL+茯苓酸组、ox-LDL+茯苓酸+inhibitor-NC组、ox-LDL+茯苓酸+miR-145-5p inhibitor组。分组处理后,CCK-8法、Edu染色检测细胞增殖;酶联免疫吸附实验(ELISA)试剂盒检测白细胞介素-1β(IL-1β)和肿瘤坏死因子α(TNF-α)水平;试剂盒检测丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)含量;实时荧光定量聚合酶链式反应(qRT-PCR)检测各组细胞miR-145-5p和KLF5 mRNA表达水平;采用流式细胞术检测细胞凋亡率;蛋白质免疫印迹法(Western Blot)检测细胞中KLF5、Bax、Bcl-2蛋白表达量。双荧光素酶报告基因实验验证miR-145-5p和KLF5的关系。结果:与对照组相比,ox-LDL组HUVEC细胞活性、增殖率、SOD和GSH、miR-145-5p表达和Bcl-2蛋白表达水平下降,IL-1β、TNF-α、MDA、凋亡率、KLF5 mRNA表达、KLF5、Bax蛋白表达水平升高(P<0.05)。与ox-LDL组相比,ox-LDL+茯苓酸组和ox-LDL+茯苓酸+inhibitor-NC组HUVEC细胞活性、增殖率、SOD和GSH、miR-145-5p表达和Bcl-2蛋白表达水平升高,IL-1β、TNF-α、MDA、凋亡率、KLF5 mRNA表达、KLF5、Bax蛋白表达水平下降(P<0.05)。与ox-LDL+茯苓酸+inhibitor-NC组相比,ox-LDL+茯苓酸+miR-145-5p inhibitor组细胞活性、增殖率、SOD和GSH、miR-145-5p表达和Bcl-2蛋白表达水平下降,IL-1β、TNF-α、MDA、凋亡率、KLF5 mRNA表达、KLF5、Bax蛋白表达水平升高(P<0.05);双荧光素酶报告基因实验验证miR-145-5p和KLF5存在靶向调控关系。结论:茯苓酸可以减少ox-LDL诱导的血管内皮细胞的凋亡,其机制可能与调节miR-145-5p/KLF5轴有关。Objective:To investigate the effect of poric acid on oxidized low densitylipoprotein(ox-LDL)-induced apoptosis of vascular endothelial cells by regulating miR-145-5p/Krüppel-like factor 5(KLF5)axis.Methods:Human umbilical vein endothelial cells(HUVEC)were grouped into control group,ox-LDL group,ox-LDL+poric acid group,ox-LDL+poric acid+inhibitor-NC group,and ox-LDL+poric acid+miR-145-5p inhibitor group.After grouping,CCK-8 method and Edu staining were applied to detect cell proliferation.The levels of interleukin(IL)-1βand tumor necrosis factor-α(TNF-α)were detected by enzyme linked immunosorbent assay(ELISA)kits.The contents of malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione(GSH)were analyzed with the kit.The expression levels of miR-145-5p and KLF5 mRNA were detected by reverse transcriptase-polymerase chain reaction(qRT-PCR).The apoptosis rate was detected by flow cytometry.Western Blot was applied to detect the expression of KLF5,Bax and Bcl-2 proteins in the cells.The relationship between miR-145-5p and KLF5 was verified by double luciferase reporter gene experiment.Results:Compared with the control group,the HUVEC cell activity,proliferation rate,SOD and GSH,the expression of miR-145-5p,and the protein expression level of Bcl-2 in ox-LDL group decreased obviously,the IL-1β,TNF-α,MDA,apoptosis rate,the expression of KLF5 mRNA,the protein expression levels of KLF5 and Bax increased obviously(P<0.05).Compared with ox-LDL group,the HUVEC cell activity,proliferation rate,SOD and GSH,the expres sion of miR-145-5p,and the protein expression level of Bcl-2 in ox-LDL+poric acid group and ox-LDL+poric acid+inhibitor-NC group increased obviously,the IL-1β,TNF-α,MDA,apoptosis rate,the expression of KLF5 mRNA,the protein expression levels of KLF5 and Bax decreased obviously(P<0.05).Compared with ox-LDL+poric acid+inhibitor-NC group,the HUVEC cell activity,proliferation rate,SOD and GSH,the expression of miR-145-5p,and the protein expression level of Bcl-2 in ox-LDL+poric acid+miR-145-5p inhibitor g

关 键 词：血管内皮细胞 细胞凋亡 茯苓酸 miR-145-5p Krüppel样转录因子5 氧化修饰的低密度脂蛋白 实验研究 

分 类 号：R285[医药卫生—中药学]