lncRNA SNHG16靶向调控miR-182-5p对缺氧/复氧诱导的心肌细胞凋亡的影响OA

Objective To explore the effect of lncRNA SNHG16 on cardiomyocyte apoptosis induced by hypoxia/reoxygenation(H/R)and its regulatory role on miR-182-5p.Methods The cells were divided into the control group(normal culture),the model group(induced by H/R),the si-NC group,the si-SNHG16 group,the si-SNHG16+anti-NC group,and the si-SNHG16+anti-miR-182-5p group.Except for the control group and the model group,the cells in the other groups were transfected with the corresponding plasmids after H/R induction.Edu assay was used to detect the cell proliferation.Flow cytometry and AO/EB staining were used to detect the cell apoptosis.ELISA was used to detect the levels of myocardial injury,inflammatory response and oxidative stress-related factors in cells.DCFDA fluorescence probe method was used to detect the production of reactive oxygen species(ROS)in cells.Western blot was used to detect the expression of apoptosis-related proteins.qRT-PCR was used to detect the levels of lncRNA SNHG16 and miR-182-5p in each group of cells.Dual-luciferase reporter gene assay and RNA pull-down assay was used to detect the targeting relationship between lncRNA SNHG16 and miR-182-5p.Results Compared with the control group,the Edu positive rate,SOD activity and miR-182-5p level in the model group decreased(P<0.05),the apoptosis rate,the proportion of apoptotic cells,the levels of lactate dehydrogenase(LDH),creatine kinase(CK),interleukin(IL)-6,IL-1β,tumor necrosis factor-alpha(TNF-α),ROS,malondialdehyde(MDA),lncRNA SNHG16,and the expressions of cleaved caspase-3(C-Caspase-3)and B-cell lymphoma-2-associated X protein(Bax)/B-cell lymphoma-2(Bcl-2)ratio increased(P<0.05).Compared with the si-NC group,the Edu positive rate,superoxide dismutase(SOD)activity and miR-182-5p level in the si-SNHG16 group increased(P<0.05),the apoptosis rate,the proportion of apoptotic cells,the levels of LDH,CK,IL-6,IL-1β,TNF-α,ROS,MDA,lncRNA SNHG16,and the expressions of C-Caspase-3 and Bax/Bcl-2 ratio decreased(P<0.05).Compared with the si-SNHG16+anti-NC group,the Edu positive rate,SOD activity and miR-182-5p level in the si-SNHG16+anti-miR-182-5p group decreased(P<0.05),the apoptosis rate,the proportion of apoptotic cells,the levels of LDH,CK,IL-6,IL-1β,TNF-α,ROS,MDA,and the expressions of C-Caspase-3 and Bax/Bcl-2 ratio increased(P<0.05).The RNA pull-down experiment revealed that there was an interaction between lncRNA SNHG16 and miR-182-5p.Compared with the cardiomyocytes co-transfected with WT-SNHG16 and miR-NC,the cardiomyocytes co-transfected with WT-SNHG16 and miR-182-5p mimics showed a relative decrease in luciferase activity(P<0.05).Conclusion Silencing lncRNA SNHG16 can reduce H/R-induced cardiomyocyte apoptosis by upregulating miR-182-5p.