Effects Of 17β-E₂ In Different Concentration On Apoptosis Of Female Rats’ Lens Epithelial Cells In Vitro Culture

objective:17β-E2 is the most active estrogen, our research aims to observe the effects of different concentration of 17β-E2 on apoptosis of lens epithelial cells induced by H2O2 in vitro culture in female rats, and investigate the regulation to expression of Bcl-2 and Bax protein, in order to approach the action and it’s mechanism of different concentration of 17β-E2 on apoptosis of lens epithelial cells, to provide the experimental evidence of estrogen prevention and treatment on cataract. Methods:Culture forty-eight diaphanous ex vivo lens of female Sprsgue-Dawley rats in medium 199. According to the difference of additional ingredient of culture fluid, we divide the forty-eight lens into six groups randomly:17β-E2+H2O2 Gruop1、17β-E2+H2O2 Group2、17β-E2+H2O2 Group3、17β-E2+H2O2 Group4、H2O2 Group and Blank Group. Use H2O2 in the final concentration of 300μmol/L to reproduce the apoptosis model of Sprsgue-Dawley rats lens epithelial cells, add 17β-E2 in different final concentration for intervention at the same time (the final concentration of 17β-E2 in 17β-E2+H2O2 Group1-4 in order:1×10-8mol/L、1×10-7mol/L 1×10-6mol/L、1×10-5mol/L). Culture the lens with incubator for twenty four hours, then compare lens opacity condition, do HE dying to lens paraffin section, and obtain anterior lens capsule and the ambitus peplos, stretch preparation, then detect the apoptosis rate by TUNEL dying, detect the expression of Bcl-2 and Bax protein by SP dying. Statistical analysis:Single factor analysis of variance, LSD method and SNK method by SPSS 13.0. Results:After cultivation (24 hours), lenses of Blank Group are completely transparent, lenses of H2O2 Group and 17β-E2 Groups are opaque, but lenses of each 17β-E2 Group are more clear than lenses of H2O2 Group(P<0.05); Apoptosis rate detected by TUNEL dying:hydrogen dioxide can successfully induce the apoptosis of lens epithelial cells, the apoptosis rate of lens epithelial cells of each 17β-E2 Groups is significantly lower than H2O2 Group’s (P<0.05), furthermore, there is an inverse correlation between the density of 17β-E2 and the apoptosis rate of lens epithelial cells (Means Plots); The expression of Bcl-2 and Bax protein detected by SP dying:in the Blank Group, both Bcl-2、Bax protein express, and the expression of Bcl-2 protein is much more powerful than the expression of Bax protein, the ratio of Bcl-2/Bax is on a high level, hydrogen dioxide can significantly down regulate the expression of Bcl-2 protein, up regulate the expression of Bax protein (P<0.05), the ratio of Bcl-2/Bax is on a low level, inversely,17β-E2 can significantly up regulate the expression of Bcl-2 protein, down regulate the expression of Bax protein(P< 0.05), furthermore, there is a positive correlation between the density of 17β-E2 and this regulation effect in a definite limit (Means Plots). Conclusion:17β-E2 can inhibit the apoptosis of female rats’lens epithelial cells induced by H2O2, and there is positive correlation between the inhibit effect and the density of 17β-E2 in a definite limit. Regulation to expression of Bcl-2 and Bax protein may be one of the molecule mechanisms.