调控circARF3/miR-338-3p对ox-LDL诱导的血管内皮细胞损伤的影响  

Effects of circARF3/miR-338-3p on ox-LDL-induced vascular endothelial cell injury

作　　者：侯永兰 金卫东 韩明磊 刘振 王建美 尚家星 HOU Yonglan;JIN Weidong;HAN Minglei;LIU Zhen;WANG Jianmei;SHANG Jiaxing(Department of Cardiovascular Medicine,Xinxiang Central Hospital(the Fourth Clinical College of Xinxiang University),Xinxiang,Henan 453000)

机构地区：[1]新乡市中心医院(新乡医学院第四临床学院)心血管内科,河南新乡453000

出　　处：《郑州大学学报（医学版）》2024年第4期509-513,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20220987)。

摘　　要：目的:探讨调控circARF3/miR-338-3p对氧化型低密度脂蛋白(ox-LDL)诱导的血管内皮细胞损伤的影响及其可能作用机制。方法:采用双荧光素酶报告实验验证circARF3与miR-338-3p的靶向关系。人脐静脉血管内皮细胞(HUVEC)分为6组:空白对照组(未处理)、模型组(50 mg/L ox-LDL处理24 h)、pcDNA组、pcDNA-circARF3组、pcDNA-circARF3+miR-NC组、pcDNA-circARF3+miR-338-3p组,后4组基于脂质体转染法分别转染相应质粒后再用ox-LDL处理24 h。采用MTT法、Annexin V-FITC/PI双染法分别检测细胞增殖及凋亡,Western blot法检测细胞中Bax、Bcl-2蛋白的表达,采用试剂盒检测培养上清液中SOD、LDH活性和MDA的水平,ELISA法检测IL-6、TNF-α水平。结果:circARF3可靶向结合、负向调控miR-338-3p。与空白对照组比较,模型组增殖抑制率、凋亡率和Bax蛋白表达均升高,而Bcl-2蛋白表达降低,培养上清液中SOD活性降低,MDA水平和LDH活性升高,IL-6、TNF-α水平升高(P<0.05);与模型组、pcDNA组相比,pcDNA-circARF3组增殖抑制率、凋亡率和Bax蛋白表达减少,Bcl-2蛋白表达提高,培养上清液中SOD活性升高,MDA水平和LDH活性降低,IL-6、TNF-α水平降低(P<0.05);与pcDNA-circARF3+miR-NC组比较,pcDNA-circARF3+miR-338-3p组增殖抑制率、凋亡率和Bax蛋白表达均升高,Bcl-2蛋白表达降低,培养上清液中SOD活性降低,MDA水平和LDH活性升高,IL-6、TNF-α水平升高(P<0.05)。结论:circARF3过表达可能通过抑制miR-338-3p削弱ox-LDL引发的血管内皮细胞损伤。Aim:To explore the effects of circARF3/miR-338-3p on oxidized low-density lipoprotein(ox-LDL)-induced vascular endothelial cell injury and its possible mechanism.Methods:The dual luciferase reporter assay was used to detect the targeting relationship between circARF3 and miR-338-3p.Human umbilical vein endothelial cells(HUVEC)were allocated into 6 groups,blank control group(not treated),model group(treated with 50 mg/L ox-LDL for 24 hours),pcDNA group,pcDNA-circARF3 group,pcDNA-circARF3+miR-NC group,and pcDNA-circARF3+miR-338-3p group,and the latter 4 groups were first transfected with the corresponding plasmids and then treated with ox-LDL for 24 hours.MTT method and AnnexinⅤ-FITC/PI were used to detect cell proliferation and apoptosis.Western blot was used to detect the expressions of Bax and Bcl-2.A kit was used to detect the activity of SOD,LDH and the level of MDA in culture supernatant.ELISA was used to detect the levels of IL-6 and TNF-αin culture supernatant.Results:circARF3 could targetedly bind and negatively regulate miR-338-3p.Compared with blank control group,the cell proliferation inhibition rate,apoptosis rate and the protein level of Bax were increased,while that of Bcl-2 decreased,the activity of SOD was decreased,the level of MDA and the activity of LDH were increased,and the levels of IL-6 and TNF-αwere increased in the model group(P<0.05).Compared with the model group and pcDNA group,the above changes in the pcDNA-circARF3 group were reversed(P<0.05).Compared with the pcDNA-circARF3+miR-NC group,the above reversed changes were partially rescued in the pcDNA-circARF3+miR-338-3p group(P<0.05).Conclusion:Overexpression of circARF3 could alleviate the injury induced by ox-LDL by inhibiting the expression of miR-338-3p.

关 键 词：人脐静脉血管内皮细胞 circARF3 miR-338-3p 细胞增殖 凋亡 氧化应激 炎症 

分 类 号：R541.4[医药卫生—心血管疾病]