邻苯二甲酸二（2-乙基）己酯诱导小鼠胚胎Leydig细胞凋亡的体外实验研究

Apoptosis of Leydig cell induced by Di（2-ethylhexyl） phthalate in vitro

目的研究邻苯二甲酸二（2-乙基）己酯EDi（2-ethylhexyl）phthalate，DEHP]对小鼠胚胎睾丸Leydig细胞凋亡诱导作用。方法DEHP50、100、200ug／ml分别作用于体外培养、纯化的胎龄16d（GD16）小鼠胚胎睾丸Leydig细胞。光镜、电镜观察Leydig细胞形态和超微结构变化，DNA末端原位标记染色法（TUNEL法）和Annexin-V／PI双染法流式细胞仪分别检测kydig细胞凋亡指数、凋亡细胞比例。结果DEHP处理组透射电镜可见凋亡典型的形态学变化，凋亡指数及凋亡细胞比例与对照组比较有显著或非常显著性差异（P〈0．05或P〈0．01）。结论DEHP能诱导小鼠胚胎睾丸Leydig细胞凋亡。

Objective To investigate the apoptosis of Leydig cells induced by Di（2-ethylhexyl） phthalate （DEHP） in vitro. Methods Testicular Leydig cells from mice （GD16） were cultured and incubated with DEHP in vitro. Cell morphological and ultrastructural changes were observed by light microscope and transmission electron microscope （TEM）. The percent of positive apoptosis Leydig cells labeling with Annexin-V/PI were analyzed by flow cytometry and the apoptosis index were detected by terminal-deoxy-nucleotidyl transferase mediated-dUTP nick end labeling （TUNEL）assay. Results Typical ultrastructural changes of apoptosis of Leydig cells were observed by TEM after DEHP treatment. Apoptotic index and percent of apoptotic cells exposed to DEHP were significantly increased compared to those in the untreated control （P〈0. 05, P〈0. 01）. Conclusions DEHP can induce embryonic Leydig cell apoptosis in vitro.