CircPTPRA通过miR-145-5p/KLF5轴调节ox-LDL诱导的血管内皮细胞凋亡的机制研究  

Mechanism of CircPTPRA Regulating ox-LDL Induced Apoptosis of Vascular Endothelial Cells Through the miR-145-5p/KLF5 Axis

作　　者：王征[1] 杨晓丽 焦清海[1] WANG Zheng;YANG Xiaoli;JIAO Qinghai(Department of Critical Care Medicine,the First Hospital of Handan,Handan 056000,China;Department of Shennei,Affiliated Hospital of Hebei University of Technology,Handan 056000,China)

机构地区：[1]邯郸市第一医院重症医学科,邯郸056000 [2]河北工程大学附属医院神内一科,邯郸056000

出　　处：《中国细胞生物学学报》2023年第10期1442-1451,共10页Chinese Journal of Cell Biology

基　　金：河北省卫健委青年科技项目(批准号:20200585)资助的课题。

摘　　要：该文旨在探讨CircPTPRA通过mi R-145-5p/KLF5轴调节氧化性低密度脂蛋白(ox-LDL)诱导的血管内皮细胞凋亡的机制。将血管内皮细胞(EVC-304)分为control组、ox-LDL组、oxLDL+si-NC组、ox-LDL+si-CircPTPRA组、ox-LDL+miR-NC组、ox-LDL+miR-145-5pmimic组、ox-LDL+si-CircPTPRA+inhibitor NC组、ox-LDL+si-CircPTPRA+miR-145-5p inhibitor组。qRT-PCR检测各组细胞中CircPTPRA、miR-145-5p和KLF5 mRNA的表达情况;MTT法、EdU染色检测细胞增殖情况;ELISA检测TNF-α、IL-6、IL-1β、MDA、SOD、GSH-Px水平;流式细胞术检测细胞凋亡率;蛋白质免疫印记法检测Ki-67、cleaved caspase-3、KLF5蛋白表达量;荧光素酶实验验证miR-145-5p与CircPTPRA、KLF5的关系。与control组相比,ox-LDL组EVC-304细胞CircPTPRA和KLF5mRNA表达量,TNF-α、IL-6、IL-1β、MDA水平,细胞凋亡率,cleaved caspase-3和KLF5蛋白水平显著升高,miR-145-5p表达量,细胞D490值(24、48 h)、增殖率,GSH-Px、SOD水平,Ki-67蛋白表达量显著降低。敲除CircPTPRA或过表达miR-145-5p都可降低细胞炎症反应程度、氧化应激水平和细胞凋亡水平。与ox-LDL+si-CircPTPRA+inhibitor NC组相比,敲低miR-145-5p可促进细胞炎症反应、氧化应激和细胞凋亡。干扰CircPTPRA可调控miR-145-5p/KLF5轴,进而减少ox-LDL诱导的血管内皮细胞凋亡。This study aims to investigate the mechanism of CircPTPRA regulating ox-LDL induced apoptosis of vascular endothelial cells through the miR-145-5p/KLF5 axis.Vascular endothelial cells(EVC304)were divided into control group,ox-LDL group,ox-LDL+si-NC group,ox-LDL+si-CircPTPRA group,oxLDL+miR-NC group,ox-LDL+miR-145-5p mimic group,ox-LDL+si-CircPTPRA+inhibitor NC group,and oxLDL+si-CircPTPRA+miR-145-5p inhibitor group.The gene expression levels of CircPTPRA,miR-145-5p,and KLF5 mRNA were detected by qRT-PCR.Cell proliferation were detected by MTT and EdU staining.The levels of TNF-α,IL-6,IL-1β,MDA,SOD and GSH-Px were detected by ELISA.Cell apoptosis rate were detected by flow cytometry.Western blot was used to detect Ki-67,cleaved caspase-3 and KLF5 protein expression levels.Luciferase assay verified the relationship between miR-145-5p and CircPTPRA and KLF5.Compared with the control group,CircPTPRA and KLF5 mRNA expression levels,TNF-α,IL-6,IL-1β,MDA levels,apoptosis rate,cleaved caspase-3 and KLF5 protein levels were significantly increased in EVC-304 cells in ox-LDL group.miR-145-5p expression,cell D490 value(24,48 h)and proliferation rate,GSH-Px and SOD levels,Ki-67 protein expression were significantly decreased.Both CircPTPRA knockout or miR-145-5p overexpression reduced cellular inflammatory response,oxidative stress and apoptosis.Compared with ox-LDL+si-CircPTPRA+inhibitor NC group,knockdown of miR-145-5p promoted cellular inflammatory response,oxidative stress and apoptosis.Interference with CircPTPRA can regulate the miR-145-5p/KLF5 axis,thereby reducing ox-LDL induced apoptosis of vascular endothelial cells.

关 键 词：CircPTPRA miR-145-5p/KLF5轴 OX-LDL 血管内皮细胞 

分 类 号：R54[医药卫生—心血管疾病]