长链非编码RNA组蛋白去甲基化酶1同系物D反义RNA1靶向微小RNA-421对过氧化氢诱导的心肌细胞氧化损伤的影响  

Effect of lncRNA JHDM1D-AS1 targeting microRNA-421 on hydrogen peroxide induced oxidative damage of cardiomyocytes

作　　者：王帅 吴申 谷阳[1] WANG Shuai;WU Shen;GU Yang(Department of Cardiovascular Medicine,Huaian First People's Hospital Affiliated to Nanjing Medical University,Huaian,Jiangsu,223300)

机构地区：[1]南京医科大学附属淮安第一医院心血管内科,江苏淮安223300

出　　处：《实用临床医药杂志》2025年第5期88-94,共7页Journal of Clinical Medicine in Practice

基　　金：江苏省淮安市自然科学研究计划项目(HAB202025)。

摘　　要：目的探讨长链非编码RNA组蛋白去甲基化酶1同系物D反义RNA1(lncRNA JHDM1D-AS1)靶向微小RNA-421(miR-421)对过氧化氢(H_(2)O_(2))诱导的心肌细胞(H9C2细胞)氧化损伤的影响。方法将H9C2细胞分为对照(Con)组、H_(2)O_(2)组、H_(2)O_(2)+pcDNA组、H_(2)O_(2)+pcDNA-JHDM1D-AS1组、H_(2)O_(2)+anti-miR-NC组、H_(2)O_(2)+anti-miR-421组、H_(2)O_(2)+pcDNA-JHDM1D-AS1+miR-NC组、H_(2)O_(2)+pcDNA-JHDM1D-AS1+miR-421组。采用实时荧光定量PCR(RT-qPCR)检测JHDM1D-AS1和miR-421表达。采用比色法测定H9C2细胞中超氧化物歧化酶(SOD)活性、丙二醛(MDA)水平以及培养液中乳酸脱氢酶(LDH)水平。采用流式细胞术评估H9C2细胞凋亡率。JHDM1D-AS1和miR-421的靶向关系通过双荧光素酶报告基因法验证。结果与Con组比较,H_(2)O_(2)组H9C2细胞JHDM1D-AS1水平、SOD活性降低,MDA水平、LDH水平、细胞凋亡率、miR-421水平升高,差异均有统计学意义(P<0.05)。与H_(2)O_(2)+pcDNA组比较,H_(2)O_(2)+pcDNA-JHDM1D-AS1组H9C2细胞SOD活性升高,miR-421表达水平、MDA水平、LDH水平、细胞凋亡率降低,差异有统计学意义(P<0.05)。与H_(2)O_(2)+anti-miR-NC组比较,H_(2)O_(2)+anti-miR-421组H9C2细胞SOD活性升高,MDA水平、LDH水平、细胞凋亡率降低,差异有统计学意义(P<0.05)。miR-421是JHDM1D-AS1的靶基因。与H_(2)O_(2)+pcDNA-JHDM1D-AS1+miR-NC组比较,H_(2)O_(2)+pcDNA-JHDM1D-AS1+miR-421组H9C2细胞SOD活性降低,MDA水平、LDH水平、细胞凋亡率升高,差异有统计学意义(P<0.05)。结论lncRNA JHDM1D-AS1通过靶向下调miR-421表达抑制细胞凋亡和氧化应激,减轻H_(2)O_(2)诱导的心肌细胞氧化损伤。Objective To investigate the effect of long non-coding RNA Jumonji C domain containing histone demethylase 1 homolog D antisense RNA1(lncRNA JHDM1D-AS1)targeting microRNA-421(miR-421)on hydrogen peroxide(H_(2)O_(2))induced oxidative damage in cardiomyocytes(H9C2 cells).Methods H9C2 cells were divided into control(Con)group,H_(2)O_(2) group,H_(2)O_(2)+pcDNA group,H_(2)O_(2)+pcDNA-JHDM1D-AS1 group,H_(2)O_(2)+anti-miR-NC group,H_(2)O_(2)+anti-miR-421 group,H_(2)O_(2)+pcDNA-JHDM1D-AS1+miR-NC group,and H_(2)O_(2)+pcDNA-JHDM1D-AS1+miR-421 group.The expressions of JHDM1D-AS1 and miR-421 were detected by real-time fluorescent quantitative PCR(RT-qPCR).The superoxide dismutase(SOD)activity,malondialdehyde(MDA)level,and lactate dehydrogenase(LDH)level in the culture medium of H9C2 cells were measured by colorimetry.Flow cytometry was used to evaluate the apoptosis rate of H9C2 cells.The targeting relationship between JHDM1D-AS1 and miR-421 was verified by dual luciferase reporter gene assay.Results Compared with the Con group,the H_(2)O_(2) group showed decreased levels of JHDM1D-AS1 and SOD activity,and increased levels of MDA,LDH,apoptosis rate,and miR-421 in H9C2 cells,with significant between-group differences(P<0.05).Compared with the H_(2)O_(2)+pcDNA group,the H_(2)O_(2)+pcDNA-JHDM1D-AS1 group exhibited increased SOD activity and decreased miR-421 expression level,MDA level,LDH level,and apoptosis rate in H9C2 cells,with significant between-group differences(P<0.05).Compared with the H_(2)O_(2)+anti-miR-NC group,the H_(2)O_(2)+anti-miR-421 group showed increased SOD activity and decreased MDA level,LDH level,and apoptosis rate in H9C2 cells,with significant between-group differences(P<0.05).MiR-421 was identified as a target gene of JHDM1D-AS1.Compared with the H_(2)O_(2)+pcDNA-JHDM1D-AS1+miR-NC group,the H_(2)O_(2)+pcDNA-JHDM1D-AS1+miR-421 group exhibited decreased SOD activity and increased MDA level,LDH level,and apoptosis rate in H9C2 cells,with significant between-group differences(P<0.05).Conclusion LncRNA JHDM

关 键 词：长链非编码RNA 组蛋白去甲基化酶1同系物D反义RNA1 微小RNA-421 过氧化氢 心肌细胞 细胞凋亡 氧化应激反应 

分 类 号：R542.2[医药卫生—心血管疾病] R346[医药卫生—内科学] Q344[医药卫生—临床医学]