Objective To explore the protective effect of geniposide on oxidative stress injury and its mechanism in H9C2.Methods The H2O2was used to construct the model of oxidative stress injury in H 9C2 cells.All the H9C2 cells were divided into three groups:PBS group, H2O2group and H2O2+geniposide group, and were stimulated by PBS, H2O2 (200 μmol/L)and H2O2+geniposide(100 μmol/L)according to the different groups.Then cells were cultured in 37℃24h.After that,the activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),and the level of malon-dialdehyde(MDA)and reactive oxygen species(ROS)were detected.Meanwhile, the mRNA levels of genes of kelch like ECH associated protein 1(KEAP 1)/NF E2 related factor 2(Nrf2)pathways and its downstream gene heme oxygenase 1 (HO-1)and SOD.Cell counting kit 8(CCK-8)and terminal deoxynucleotidyl transferase mediated dUTP biotin nick end la-beling assay(TUNEL)were used respectively to detect the viability and apoptosis of H 9C2.Results The activities of SOD and GSH Px in H9C2 cells were decreased after H2O2,and geniposide could increase the decreased SOD and GSH-Px.Geni-poside also inhibited the production of MDA induced by H 2O2.ROS production was also suppressed by the treatment of geni-poside.In the H2O2treatment group,the mRNA level of KEAP1 was increased and the mRNA level of Nrf 2,HO-1 and SOD-1 were decreased.However,in H2O2plus geniposide group,these pathological changes have been reversed.Meanwhile,the use of geniposide increased cell viability and decreased the apoptosis of H 9C2 cells.Conclusion Geniposide can inhibit the oxidative injury induced by H 2O2via the activation of Keap1/Nrf2 pathway in H9C2 cells.%目的 探讨栀子苷(GE)对H9C2心肌细胞氧化应激损伤的保护作用及机制.方法 研究于2016年12月—2017年9月在武汉大学心血管病研究所进行.采用过氧化氢(H2O2)建立H9C2细胞氧化应激损伤模型.将细胞分为3组:PBS组、H2O2组以及H2O2+GE组,分别给予磷酸缓冲盐溶液(PBS)、H2O2(200μmol/L)、H2O2+GE(100μmol/L),刺激后继续培养24h.检测各组内超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性,丙二醛(MDA)、活性氧簇(ROS)水平,检测Kelch样环氧氯丙烷相关蛋白-1(KEAP1)/核因子E2相关因子2(Nrf2)通路及其下游的血红素加氧酶-1(HO-1)和SOD-1的mRNA水平,CCK-8检测细胞存活率,原位末端缺口标记法(TUNEL)检测细胞凋亡.结果h2O2处理后,H9C2心肌细胞内SOD和GSH-Px的活性下降,与H2O2组比较,H2O2+GE组细胞SOD和GSH-Px活性增加;H2O2处理可导致H9C2心肌细胞MDA水平明显升高,栀子苷可抑制这种脂质过氧化产物的产生;H2O2可诱导H9c2细胞ROS的产生,栀子苷处理后ROS的水平显著下降.H2O2刺激细胞后,KEAP1转录水平上升,Nrf2、HO-1、SOD-1转录水平下降;栀子苷处理后,KEAP1转录水平下降、Nrf2、HO-1、SOD-1转录水平回升;栀子苷明显升高细胞存活率,减轻细胞凋亡.结论 栀子苷可通过激活KEAP1/Nrf2通路抑制H2O2引起的心肌细胞的氧化损伤.
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