RESUMO
Objective To evaluate the effect of oxidative injury induced by peroxide oxidase on Klotho expression in mouse renal tubular epithelial cells (TCMK-1) and to explore the possible pathway.Methods TCMK-1 cells were exposed to H2O2 of different concentrations.Reactive oxygen species (ROS) was examined byflow cytometrry.Cell viability was assessed by CCK-8.Cell apoptosis was evaluated by flow cytometry and Hoechst 33258 staining.The expression of Klotho,apoptosis-associated proteins and anti-oxidant enzymes were determined by Western blotting.Results Compared with control group,after H2O2 stimulating TCMK-1 cell,ROS was dramatically elevated (all P ＜ 0.05) and the expression of anti-oxidant enzymes,SOD2 and CAT went down (all P ＜ 0.05);the expression of Klotho was inhibited (all P ＜ 0.05);cell viability of TCMK-1 cells was decreased (all P ＜ 0.05) in a dose-dependent manner (0.3 to 0.9 mmol/L);cell apoptosis was significantly increased in TCMK-1 cells following the concentration of H2O2 (all P ＜ 0.05);Bax/Bcl-2 and the phosphororation of JNK and p38 were obviously elevated in TCMK-1 by H2O2 induction (all P ＜ 0.05).Conclusion Oxidative injuries induced by H2O2 significantly suppresses the expression of Klotho in TCMK-1 cells.And cell apoptosis was increased,p38 and JNK pathway was activated.
