Effect of Up-regulated lncRNA MEG3 on Hypoxia/Reoxygenation Injury of Rats Myocardial Cell Lines
Objective:To explore the protective effect of maternal expression of long non-coding RNA(lncRNA)maternally expressed gene 3(MEG3)on hypoxia/reoxygenation(H/R)in rat myocardial H9c2 cells and its relationship with the nuclear factor E2 related factor 2(Nrf2)/antioxidant response element(ARE)pathway.Methods:H9c2 cells were divided into control group,H/R group,H/R+Ad-Green fluorescent protein(GFP)group,H/R+Ad-MEG3 group,H/R+Ad-MEG3+si-NC group and H/R+Ad-MEG3+si-Nrf2 group.Real-time fluorescent quantitative reverse transcriptase polymerase chain reaction(RT-qPCR)was used to detect MEG3 expression.Methyl thiazolyl tetrazolium(MTT)method was used to detect the viability of cells.Flow cytometry was used to detect the apoptosis rate.2',7'-dichlorofluorescent yellow diacetate(DCFH-DA)method was used to detect the level of reactive oxygen species(ROS).Enzyme-linked immunosorbent assay(ELISA)was used to detect the activities of superoxide dismutase(SOD)and catalase(CAT).Western Blot was used to detect the expression levels of Nrf2/ARE pathway related proteins.Results:Compared with the control group,the H9c2 cell apoptosis rate,cell ROS level and Nrf2,heme oxidase 1(HO-1)and quinone oxidoreductase-1(NQO-1)protein levels increased in the H/R group,while the cell viability,the MEG3 expression level in the cells,SOD and CAT activities reduced(P<0.05).MEG3 overexpression partially reversed the effects of H/R on H9c2 cells and Nrf2/ARE pathway proteins(P<0.05).Knockdown of Nrf2 could weaken the protective effect of MEG3 overexpression on H/R injury(P<0.05).Conclusion:Up-regulating the expression of MEG3 could inhibit oxidative stress and apoptosis to protect H/R myocardial Injury,which may be related to the activation of Nrf2/ARE antioxidant signals.
hypoxia/reoxygenationlong non-coding RNA maternally expressed gene 3cardiomyocyte H9c2experimental study
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