摘要
目的 探讨长链非编码RNA(lncRNA)NUTM2A-AS1对氧化低密度脂蛋白(oxLDL)诱导的血管内皮细胞损伤的影响及分子机制。方法 将人脐静脉血管内皮细胞(HUVEC)在DMEM培养基中培养,采用100μg/mL oxLDL处理的HUVEC纳入oxLDL组,常规培养的细胞纳入Con组。将lncRNA NUTM2A-AS1干扰表达载体及阴性对照、miR-129-5p模拟物及阴性对照转染至HUVEC后采用100μg/mL oxLDL处理后的细胞分别纳入oxLDL+si-NUTM2A-AS1组、oxLDL+si-NC组、oxLDL+miR-129-5p组、oxLDL+miR-NC组。将lncRNA NUTM2A-AS1干扰表达载体与微小RNA(miR)-129-5p抑制剂或阴性对照共转染至HUVEC后采用100μg/mL的oxLDL处理的细胞分别纳入oxLDL+si-NUTM2A-AS1+anti-miR-129-5p组、oxLDL+si-NUTM2A-AS1+anti-miR-NC组。采用试剂盒测量细胞中丙二醛(MDA)含量、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性;采用流式细胞术检测细胞凋亡;采用蛋白质印迹法检测蛋白表达;采用双荧光素酶报告实验及RNA下拉实验检测NUTM2A-AS1与miR-129-5p的靶向关系。结果 与Con组比较,oxLDL组lncRNA NUTM2A-AS1表达水平升高,miR-129-5p表达水平降低,MDA含量升高,SOD、GSH-Px活性降低,血管内皮细胞凋亡率和cleaved-caspase3、cleaved-caspase9表达水平升高,差异有统计学意义(P<0.05)。干扰lncRNA NUTM2A-AS1表达或过表达miR-129-5p后,MDA含量降低,SOD、GSH-Px活性升高,细胞凋亡率降低,差异有统计学意义(P<0.05)。lncRNA NUTM2A-AS1敲低介导的oxLDL条件下血管内皮细胞的损伤抑制可以被miR-129-5p下调逆转。lncRNA NUTM2A-AS1靶向调控miR-129-5p。结论 干扰lncRNA NUTM2A-AS1表达通过靶向上调miR-129-5p抑制oxLDL诱导的血管内皮细胞损伤。
Objective To explore the effect of long non-coding RNA(lncRNA) NUTM2A-AS1 on the damage of vascular endothelial cells induced by oxidized low density lipoprotein(oxLDL) and its molecular mechanism.Methods Human umbilical vein endothelial cells(HUVECs) were cultured in DMEM medium.The HUVECs treated with 100 μg/mL oxLDL were assigned to oxLDL group,while those cultured under normal conditions were assigned to Con group.After transfection of the lncRNA NUTM2A-AS1 interference expression vector and negative control,microRNA-129-5p mimic and negative control into HUVECs,the cells treated with 100 μg/mL oxLDL were assigned to oxLDL+si-NUTM2A-AS1 group,oxLDL+si-NC group,oxLDL+miR-129-5p group,and oxLDL+miR-NC group,respectively.After co-transfection of the lncRNA NUTM2A-AS1 interference expression vector and miR-129-5p inhibitor or negative control into HUVECs,the cells treated with 100 μg/mL oxLDL were assigned to oxLDL + si-NUTM2A-AS1 + anti-miR-129-5p group and oxLDL + si-NUTM2A-AS1 +anti-miR-NC group.The levels of malondialdehyde(MDA) in cells,as well as the activities of superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) were measured using kits.Cell apoptosis was detected by flow cytometry.Protein expression was detected by Western blot.The targeting relationship between NUTM2A-AS1 and miR-129-5p was detected by dual luciferase reporter assay and RNA pull-down experiments.Results Compared with the Con group,the expression level of lncRNA NUTM2A-AS1 was increased,the expression level of miR-129-5p was decreased,the content of MDA was increased,the activities of SOD and GSH-Px were decreased,the apoptosis rate of vascular endothelial cells and the expression levels of cleaved-caspase3 and cleaved-caspase9 were increased in the oxLDL group(P< 0.05).After interference of lncRNA NUTM2A-AS1 expression or overexpression of miR-129-5p,the content of MDA was decreased,the activities of SOD and GSH-Px were increased,cell apoptosis was reduced(P< 0.05).The damage to vascular endothelial cells mediated by knockdown of lncRNA NUTM2A-AS1 under oxLDL conditions can be reversed bCyo ndcolwunsiroengulation of miR-129-5p.LncRNA NUTM2A-AS1 targeted miR-129-5p for regulation. Interference of lncRNA NUTM2A-AS1 expression can inhibit oxLDL-induced vascular endothelial cell damage by targeting miR-129-5p for upregulation.
作者
李晓宇
张永咏
秦娟
杨忠信
LI Xiaoyu;ZHANG Yongyong;QIN Juan;YANG Zhongxin(Department of Cardiovascular Medicine,the First Affiliated Hospital of Henan University,Kaifeng,Henan,475000;Department of Thoracic Surgery,the First Affiliated Hospital of Henan University,Kaifeng,Henan,475000)
出处
《实用临床医药杂志》
CAS
2024年第3期45-50,共6页
Journal of Clinical Medicine in Practice
基金
河南省医学科技攻关计划联合共建项目(LHGJ20190501)。
