己烯雌酚对体外培养仓鼠生精细胞毒性作用的研究

Study on Toxic Effects of Diethylstilbestrol on Cultured Hamster Spermatogenic Cells

为研究己烯雌酚(DES)对仓鼠生精细胞毒性作用机制,试验采用原代培养的仓鼠睾丸生精细胞,用10、30、90μmol/L DES处理,倒置显微镜观察生精细胞生长状况,MTT法测定处理后8 h生精细胞存活率,分光光度法检测细胞培养液中超氧化物歧化酶(SOD)、丙二醛(MDA)、细胞中谷胱甘肽过氧化物酶(GSH-Px)的含量。结果贴壁培养的生精细胞用DES处理后,细胞活力显著降低,与正常对照组比较差异极显著(P<0.01),随着DES剂量的增加,细胞中SOD、GSH-Px含量显著下降,MDA显著上升。说明DES对体外培养生殖细胞毒性作用与ROS密切相关,DES通过降低抗氧化酶水平,增加ROS含量,干扰生精细胞正常功能,表明氧化损伤可能是环境雌激素生殖毒性的作用机制之一。

To explore the mechanism on diethylstilbestrol（DES） induced toxic effects on cultured hamster spermatogenic cells,the spermatogenic cells of hamster were treated with different concentrations of DES.The growing status of cultured cells was observed under the inverted microscope.The viability of spermatogenic cells were measured by MTT.The contents of superoxide dismutase（SOD） and malondialdehyde（MDA） in supernatants and glutathione peroxidase（GSH-Px） in cells were detected by spectrophotography.The results showed that exposure to DES,there is a significant reduction in cell viability compared with the control group（P〈0.01）.As well as the increase of DES,the contents of SOD and GSH-Px all decreased which led to the increase in MDA.It is concluded that there is a close relationship between reproductive toxicity of DES with reactive oxygen species（ROS）.DES impairs the normal function of spermatogenic cells through decreasing antioxidase avtivity,resulting in the increases of ROS.These results indicated that oxidative damage may be one of the mechanisms of reproductive toxicity and cellular apoptosis induced by environmental estrogen.