ABSTRACT
Objective To investigate the protective effects of 17β-estradiol on human lens epithelial (HLE) cells in oxidative damage induced by H2 O2 and its involved mechanisms.Methods HLE cells cultured in vitro were collected and divided into 4 groups;cells in negative control group were cultured with normal medium,cells with 80％ fusion in n2O2 damage group was treated with 100 μmol · L-1 H2O2 for 12 h,cells in 17β-estradiol low dose group were incubated with 10 μmol · L-1 17β-estradiol for 24 h then subjected to 100 μmol · L-1 H2O2 for 12 h and ceEs in 17β-estradiol high dose group were cultured in 100 μmol · L-1 17β-estradiol for 24 h then treated with 100 μrnol · L-1 H2O2 for 12h.Next,cell viability was tested by CCK-8 colorimetric assay,while apoptotic rate was detected by flow cytometry,and intracellular reactive oxygen species (ROS) level was detected by H2 DCFDA fluorescence probe labeling method,as well as the contents of catalase (CAT),superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined by 721 D spectrophotometer.Results When compared with the negative control group,the survival rate of HLE ceils in the H2 O2 damage group was significantly decreased,and there was significantly different between both groups (P ＜0.05).Moreover,the survival rate of the 17β-estradiol low dose group (67.44％) and high dose group (78.52％) was obviously higher than that of the H2O2 damage group (59.34％),and the difference was statistically significant (P ＜ 0.05).After H2O2-induced injury,there was significant difference in the apoptotic rate of HLE cells between the H2O2 damage group (41.30 ±3.21)％ and the negative control group (1.67+0.32)％.In addition,the apoptotic rates of the 17β-estradiol low dose group and high dose group was (20.97 + 1.13)％ and (14.27 + 0.90)％ respectively,which was statistically different from the H2 O2 damage group (all P ＜ 0.05).H2 DCFDA fluorescent labeling test results showed that ROS fluorescence signal intensity gradually weakened after treated with 17β-estradiol.Besides,17β-estradiol significantly increased the expression of CAT、SOD and GSH-Px in HLE cells.Conclusion 17β-estradiol has obvious protective effects on HLE cells from the damage induced by H2O2.
ABSTRACT
Objective To investigate the protective effects of 17β-estradiol on human lens epithelial (HLE) cells in oxidative damage induced by H2 O2 and its involved mechanisms.Methods HLE cells cultured in vitro were collected and divided into 4 groups;cells in negative control group were cultured with normal medium,cells with 80％ fusion in n2O2 damage group was treated with 100 μmol · L-1 H2O2 for 12 h,cells in 17β-estradiol low dose group were incubated with 10 μmol · L-1 17β-estradiol for 24 h then subjected to 100 μmol · L-1 H2O2 for 12 h and ceEs in 17β-estradiol high dose group were cultured in 100 μmol · L-1 17β-estradiol for 24 h then treated with 100 μrnol · L-1 H2O2 for 12h.Next,cell viability was tested by CCK-8 colorimetric assay,while apoptotic rate was detected by flow cytometry,and intracellular reactive oxygen species (ROS) level was detected by H2 DCFDA fluorescence probe labeling method,as well as the contents of catalase (CAT),superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined by 721 D spectrophotometer.Results When compared with the negative control group,the survival rate of HLE ceils in the H2 O2 damage group was significantly decreased,and there was significantly different between both groups (P ＜0.05).Moreover,the survival rate of the 17β-estradiol low dose group (67.44％) and high dose group (78.52％) was obviously higher than that of the H2O2 damage group (59.34％),and the difference was statistically significant (P ＜ 0.05).After H2O2-induced injury,there was significant difference in the apoptotic rate of HLE cells between the H2O2 damage group (41.30 ±3.21)％ and the negative control group (1.67+0.32)％.In addition,the apoptotic rates of the 17β-estradiol low dose group and high dose group was (20.97 + 1.13)％ and (14.27 + 0.90)％ respectively,which was statistically different from the H2 O2 damage group (all P ＜ 0.05).H2 DCFDA fluorescent labeling test results showed that ROS fluorescence signal intensity gradually weakened after treated with 17β-estradiol.Besides,17β-estradiol significantly increased the expression of CAT、SOD and GSH-Px in HLE cells.Conclusion 17β-estradiol has obvious protective effects on HLE cells from the damage induced by H2O2.