lncRNA PVT1通过靶向miR-761对缺氧复氧诱导的心肌细胞损伤的影响  

Effect of lncRNA PVT1 on cardiomyocyte damage induced by hypoxia/reoxygenation by targeting miR-761

作　　者：黄柳[1] 崔坤 贾妍[1] 郭炳彦[1] 李拥军 HUANG Liu;CUI Kun;JIA Yan;GUO Bingyan;LI Yongjun(Department of Cardiology,the Second Hospital of Hebei Medical University,Shijiazhuang,Hebei 050000,China)

机构地区：[1]河北医科大学第二医院心血管内科,河北省石家庄市050000

出　　处：《中国动脉硬化杂志》2021年第4期295-300,共6页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金项目(81570345);河北省卫生健康委员会课题(20190501)。

摘　　要：目的探讨lncRNA PVT1对缺氧复氧(H/R)诱导的心肌细胞凋亡、氧化应激的影响及其对miR-761的调控作用。方法采用H/R诱导大鼠心肌细胞H9c2建立细胞损伤模型,分别将si-NC、si-PVT1、miR-mimics-NC、miR-761 mimics、si-PVT1与miR-inhibitor-NC、si-PVT1与miR-761 inhibitor转染至H/R诱导的心肌细胞;采用qRT-PCR法检测PVT1、miR-761的表达量;采用流式细胞术检测细胞凋亡率;采用试剂盒检测丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的含量;双荧光素酶报告实验检测PVT1与miR-761的靶向关系;Western blot法检测B淋巴细胞瘤2(Bcl-2)、B淋巴细胞瘤2相关蛋白(Bax)的蛋白表达量。结果与对照组比较,H/R组PVT1的表达水平升高(P<0.05),miR-761的表达水平降低(P<0.05)。与H/R+si-NC组比较,H/R+si-PVT1组凋亡率及Bax蛋白水平降低(P<0.05),Bcl-2蛋白水平及SOD、CAT的活性升高(P<0.05),MDA的含量降低(P<0.05)。与H/R+miR-mimics-NC组比较,H/R+miR-761 mimics组凋亡率及Bax蛋白水平降低(P<0.05),Bcl-2蛋白水平及SOD、CAT的活性升高(P<0.05),MDA的含量降低(P<0.05)。双荧光素酶报告实验证实PVT1可靶向结合miR-761;干扰miR-761可明显逆转沉默PVT1对H/R诱导的心肌细胞氧化应激及凋亡的作用。结论沉默PVT1可通过上调miR-761而抑制细胞凋亡及氧化应激从而减轻H/R诱导的心肌细胞损伤。Aim To explore the effect of lncRNA PVT1 on oxidative stress and cardiomyocyte apoptosis induced by hypoxia/reoxygenation(H/R)and its regulatory effect on miR-761.Methods Rat cardiomyocyte H9c2 cells were treated with hypoxia and reoxygenation(H/R)to establish a cell injury model.si-NC,si-PVT1,miR-mimics-NC,miR-761 mimics,si-PVT1 and miR-inhibitor-NC,si-PVT1 and miR-761 inhibitor were transfected into H/R-induced cardiomyocytes.qRT-PCR method was used to detect the expression of PVT1 and miR-761.Flow cytometry was used to detect the apoptosis rate.The levels of malondialedhyde(MDA),superoxide dismutase(SOD),and catalase(CAT)were detected by Kits.The dual luciferase reporter experiment was used to analyze the targeting relationship between PVT1 and miR-761.Western blot was used to detect the expression of Bcl-2 and Bax protein.Results Compared with the control group,the expression level of PVT1 in the H/R group was increased(P<0.05),and the expression level of miR-761 was decreased(P<0.05).Compared with the H/R+si-NC group,the apoptosis rate and the protein level of Bax were decreased in the H/R+si-PVT1 group(P<0.05),the protein level of Bcl-2 and the activities of SOD and CAT were increased(P<0.05),and the content of MDA was decreased(P<0.05).Compared with H/R+miR-mimics-NC group,the apoptosis rate and the protein level of Bax were decreased in H/R+miR-761 mimics group(P<0.05),the protein level of Bcl-2 and the activities of SOD and CAT were increased(P<0.05),the content of MDA was decreased(P<0.05).The result of dual luciferase report experiment confirmed that miR-761 was the downstream target of PVT1.Interference with miR-761 could obviously reverse the effect of silencing PVT1 on H/R-induced oxidative stress and apoptosis of cardiomyocytes.Conclusion Silencing PVT1 could inhibit H9c2 cell apoptosis and oxidative stress by up-regulating miR-761,thereby reducing H/R-induced cardiomyocyte damage.

关 键 词：lncRNA PVT1 miR-761 缺氧复氧 心肌细胞 细胞凋亡 氧化应激 

分 类 号：R5[医药卫生—内科学]