邻苯二甲酸二乙基己酯对小鼠胚胎Leydig细胞毒性作用研究

目的探讨邻苯二甲酸二乙基己酯(DEHP)对小鼠胚胎睾丸问质细胞(Leydig细胞)的毒性作用。方法小鼠胚胎Leydig细胞体外原代培养。观察DEHP导致的Leydig细胞形态及超微结构改变,用MTT法和Annexin-V／PI流式细胞术分别检测DEHP在培养基内终浓度为25、510、100和200mg／L时对Leydig细胞活力影响及毒性作用。结果Leydig细胞活力随着DEHP浓度增加和作用时间延长而明显下降。Annexin-V／PI流式细胞术检测提示:死亡细胞比例随DEHP浓度升高和作用时间延长而显著增加,同时细胞增殖速度变慢,甚至增殖停止,伴随一系列细胞形态学及超微结构改变,呈剂量、时间依赖关系。结论DEHP能降低Leydig细胞的活性并促进其死亡,有明显细胞毒性。

Cytotoxicity of Di (2-ethylhexyl) phthalate on Leydig cells primary cultured from the fetal mice

Objective To detect the toxic effects of Di (2-ethylhexyl) phthalate (DEHP) on Leydig cells primarily cultured from the fetal mice testis. Methods The Leydig cells were isolated from fetal mice testis and were cultured in DMEM/F12 medium different conceutration. The viability of Leydig cells incubated with DEHP at different concentration was measured using MTT assay and toxic effect were evaluated by flow cytometric assay. The cell morphological and ultrastructure alteration were observed during various culture time and DEHP doses. Results Leydig cell viability decreased significantly in a manner of dose-dependent and time-dependent while treated with DEHP. Annexin- V /PI flow cytometric assay revealed that the number of dead Leydig cells grew up along with exposing to DEHP concentration increasing. The Leydig cell morphology and ultrstructure were affected significantly by DEHP concentration and doses. Conclusion DEHP had the cytotoxic effects that affected the viability of fetal mice Leydig cells and increase the percent of dead cells in vitro.