草苁蓉多糖调控miR-138-5p对H_(2)O_(2)诱导H9c2心肌细胞凋亡和氧化损伤的影响

Effects of Boschniakia Rossica Polysaccharides on H_(2)O_(2)-Induced Apoptosis and Oxidative Stress Damage in H9c2 Cardiomyocyte by Regulating miR-138-5p

目的探讨草苁蓉多糖对H_(2)O_(2)诱导H9c2心肌细胞凋亡、氧化损伤的影响和机制。方法使用50μmol/L H_(2)O_(2)孵育H9c2心肌细胞2 h建立细胞损伤模型。将H9c2心肌细胞分为con组、H_(2)O_(2)组、H_(2)O_(2)+草苁蓉多糖-L组、H_(2)O_(2)+草苁蓉多糖-M组、H_(2)O_(2)+草苁蓉多糖-H组、H_(2)O_(2)+miR-NC组、H_(2)O_(2)+miR-138-5p组、H_(2)O_(2)+anti-miR-138-5p组、H_(2)O_(2)+草苁蓉多糖+anti-miR-138-5p组。通过细胞计数试剂盒8(CCK-8)、流式细胞测量术分析细胞活力和凋亡率;测定超氧化物歧化酶(SOD)活性、丙二醛(MDA)水平及乳酸脱氢酶(LDH)释放量;实时荧光定量聚合酶链式反应(RT-qPCR)分析miR-138-5p表达水平。结果与con组比较,H_(2)O_(2)组细胞活力、SOD活性、miR-138-5p表达降低(P<0.05),凋亡率、MDA水平、LDH释放量升高(P<0.05)。与H_(2)O_(2)组比较,H_(2)O_(2)+草苁蓉多糖-L组、H_(2)O_(2)+草苁蓉多糖-M组、H_(2)O_(2)+草苁蓉多糖-H组细胞活力、SOD活性、miR-138-5p表达升高(P<0.05),凋亡率、MDA水平、LDH释放量降低(P<0.05)。与H_(2)O_(2)组、H_(2)O_(2)+miR-NC组比较,H_(2)O_(2)+miR-138-5p组细胞活力、SOD活性升高(P<0.05),凋亡率、MDA水平、LDH释放量降低(P<0.05)。与H_(2)O_(2)组比较,H_(2)O_(2)+anti-miR-138-5p组细胞活力、SOD活性降低(P<0.05),凋亡率、MDA水平、LDH释放量升高(P<0.05)。与H_(2)O_(2)+草苁蓉多糖组比较,H_(2)O_(2)+草苁蓉多糖+anti-miR-138-5p组细胞活力、SOD活性降低(P<0.05),凋亡率、MDA水平、LDH释放量升高(P<0.05)。结论草苁蓉多糖通过上调miR-138-5p抑制氧化应激和细胞凋亡,保护H9c2心肌细胞免受H_(2)O_(2)诱导的损伤。

Objective To investigate the effect of Boschniakia rossica polysaccharide on H_(2)O_(2)-induced cardiomyocyte H9c2 apoptosis and oxidative damage,and its mechanism.Methods H9c2 cardiomyocytes were incubated with 50μmol/L H_(2)O_(2)for 2 h to establish a cell injury model.H9c2 cardiomyocytes were divided into control(con)group,H_(2)O_(2)group,H_(2)O_(2)+Boschniakia rossica polysaccharide-L group,H_(2)O_(2)+Boschniakia rossica polysaccharide-M group,H_(2)O_(2)+Boschniakia rossica polysaccharide-H group,H_(2)O_(2)+miR-NC group,H_(2)O_(2)+miR-138-5p group,H_(2)O_(2)+anti-miR-138-5p group,H_(2)O_(2)+Boschniakia rossica polysaccharide+anti-miR-138-5p group.Cell survival rate and apoptosis rate were detected by cell counting kit 8(CCK-8)and flow cytometry.Superoxide dismutase(SOD)activity,malondialdehyde(MDA)level,and lactate dehydrogenase(LDH)were analyzed.miR-138-5p expression was detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results Compared with the con group,the cell viability,SOD activity,and miR-138-5p expression in the H_(2)O_(2)group were decreased(P<0.05),while the apoptosis rate,MDA level,and LDH release were increased(P<0.05).Compared with the H_(2)O_(2)group,the cell viability,SOD activity,and miR-138-5p expression in the H_(2)O_(2)+Boschniakia rossica polysaccharide-L group,H_(2)O_(2)Boschniakia rossica polysaccharide-M group,and H_(2)O_(2)Boschniakia rossica polysaccharide-H group were increased,apoptosis rate,while MDA level,and LDH release were decreased(P<0.05).Compared with the H_(2)O_(2)group and the H_(2)O_(2)+miR-NC group,the cell viability and SOD activity in the H_(2)O_(2)+miR-138-5p group were increased(P<0.05),while the apoptosis rate,MDA level,and LDH release were decreased(P<0.05).Compared with the H_(2)O_(2)group,the cell viability and SOD activity in the H_(2)O_(2)+anti-miR-138-5p group were decreased(P<0.05),while the apoptosis rate,MDA level,and LDH release were increased(P<0.05).Compared with the H_(2)O_(2)+Boschniakia rossica polysaccharide group,the cell viability and SOD activity in the H_(2)O_(2)+Boschniakia rossica polysaccharide+anti-miR-138-5p group were decreased(P<0.05),while apoptosis rate,MDA level,and LDH release were increased(P<0.05).Conclusion Boschniakia rossica polysaccharide inhibited oxidative stress and apoptosis by upregulating miR-138-5p,and protected H9c2 cardiomyocyte from H_(2)O_(2)-induced damage.