Mechanisms of Gancao Xiexin Decoction on improving oxidative stress injury of HT29 cells by regulating Keap1-Nrf2 signaling pathway
Objective To investigate the mechanism of Gancao Xiexin Decoction(GXD)in improving oxidative stress(OS)damage of HT29 cells by regulating Keap1-Nrf2 signaling pathway.Methods UHPLC-Q-TOF-MS high-resolution LC/MS approach was ap-plied to identify compounds in GXD.HT29 cells were induced by H2O2 to establish OS cell model.We divided cells into normal control group,stress model group,GXD low-and high-dose group;we assessed their survival rate,apoptosis rate,cycle distribution,and the expressions of apoptosis-related proteins,to verify the protective effect of GXD.We divided cells into si-NC group,si-NC+stress group,si-NC+stress+GXD group and si-Nrf2+stress+GXD group;we detected the levels of ROS,MDA,SOD and T-AOC,the expression and distribution of proteins in Keap1-Nrf2 signaling pathway,to clarify the mechanism of Nrf2-ARE signaling pathway in reducing OS damage of cells by GXD.Results A total of 20523 compounds were identified in GXD,including 10 main com-pounds.Compared with stress model group,GXD treatment increased the survival rates and the proportions of cells in G2 phase(P＜0.05),as well as the expressions of Bcl-2(P＜0.01),while decreased the apoptosis rates(P＜0.05)and the expressions of Bax and cleaved-caspase-3(P＜0.05).Compared with si-NC+stress group,the levels of SOD,T-AOC,total Nrf2,Nrf2,HO-1 and NQO1 in si-NC+stress+GXD group were increased(P＜0.01),the levels of ROS,MDA and Keap1 were decreased(P＜0.01).Compared with si-NC+stress+GXD group,the above indexes in the si-Nrf2+stress+GXD group showed an opposite trend.Conclusion By activating Keap1-Nrf2 pathway signal transduction,GXD restores the oxidation-antioxidant system balance and allevi-ates cell injury and apoptosis,which may be the mechanism of its ameliorating OS injury.
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