Study On Effect And Mechanisms Of Genistein On Human Lens Epithelial Cell Injured By H₂O₂

ObjectiveTo investigate the effect of Chinese medicine with estrogen activity, genistein(Gen), on human lens epithelial cell(HLEC) injured by H2O2,to reveal the cellular and molecular mechanism of preventing and treating cataract by phytoestrogen in the fields of oxidation and apoptosis, in order to provide scientific evidence for pursue effective midicine to prevent and treat cataract.MethodHuman lens epithelia cell HLEC were cultured with Gen and E2respectively,then, the cells were cultured with H2O2. And then, the damaged HLEC were used for following studies:1. The effect of different concentrations of Gen on activity of HLEC damaged by H2O2were detected by methyl thiazolyl tetrazolium (MTT).2. The effect of Gen on oxidative damage of HLEC injured by H2O2:the activities of superoxide dismutase(SOD) and catalase(CAT), the content of glutathione(GSH) and malondialdehyde (MDA) of HLEC damaged by H2O2were measured via chemical chromatometry to study the anti-oxidative effects of Gen and the enzymatic mechanism.3. The effect of Gen on apoptosis of HLEC injured by H2O2(1) The effect of Gen on morphology of HLEC damaged by H2O2was observed under transmission electron-microscope.(2) The effect of Gen on apoptotic rate of HLEC damaged by H2O2was analyzedusing flow cytometry.(3) The effect of Gen on mitochondrial transmembrane potential ATM) of HLEC damaged by H2O2was analyzed using flow cytometryResults1. The result of MTT test show that the absorbance values(A) of H2O2group was lower obviously than that in control group(P<0.01).The A values of HLEC in different group of Gen were increased obviously to compare with H2O2group. 2. The effect of Gen on oxidative damage of HLEC injured by H2O2:the result of chemical test show that the activities or contents of SOD,CAT and GSH decreased in H2O2group, however, in E2group,the activities of SOD and CAT,increased markedly compared with H2O2group(P<0.01),the content of GSH also increased obviously(P<0.05). In Gen group, the activity of CAT and the content of GSH increased markedly compared with H2O2group(P<0.05), the activity of SOD also increased obviously(P<0.01). The MDA contents increased in H2O2group,while compared with H2O2group,it decreased markedly in E2group (P<0.05),and it also decreased markedly in Gen group(P<0.01).3. The effect of Gen on apoptosis of HLEC injured by H2O2.(1) The morphological changes of apoptosis by transmission electron microscopy showed that the typical morphological changes of apoptosis were observed under transmission electronmicroscope,such as heterochromatin condensation and aggregated at the nuclear periphery,and nuclear fragmentation as well in HLEC induced by H2O2.However,E2and Gen group can alleviated these changes of morphology.(2) The result of flow cytometry showed that the apoptotic rate of HLEC increased in H2O2group comparing with control group,and the apoptotic rate decreased markedly in E2and Gen group comparing with H2O2group (P<0.01).(3) The result of flow cytometry showed that mitochondrial transmembrane potential (ΔΨM) in H2O2group reduced obviously compared to control group(P<0.01)and ΔΨM in E2and Gen group is increased remarkably compared to H2O2group(P<0.01).Conclusion1.H2O2can induce oxidative damages of HLEC. E2can safeguard oxidative damages injured by H2O2, Gen also can safeguard oxidative damages injured by H2O2, and show estrogen-like effect,2.1ncreasing the SOD,CAT and GSH activity or content and decreasing the MDA content may be the important mechanism of Gen preventing and treating injury of HLEC damaged by H2O2.3.H2O2can induce apoptosis of HLEC. E2can reduced apoptosis of HLEC injured by H2O2.Gen also can reduced apoptosis of HLEC injured by H2O2, and show estrogen-like effect.4.Decreasing apoptosis rate and rising level of ΔΨM may be the mechanism of Gen preventing and treating injury of HLEC damaged by H2O2.5.These conclusions show that as one of phytoestrogens,Gen may show estrogen-like effect. Through increasing antioxidant capacity and decreasing apoptosis degree,Gen show protective effect to HLEC injured by H2O2.