DEHP体外影响大鼠睾丸间质细胞Bax和Bcl-2基因表达的实验研究

摘要

Objective To investigate the effects of di-2-ethylhexyl phthalate ( DEHP) on the expressions of Bax and Bcl-2 genes, and on the apoptosis in rat Leydig cells in vitro. Methods The rat Leydig cells were separated and cultured, and the Leydig cells were treated with different concentrations of DEHP ( 10,50,100nmol/L )for 24h. Real time PCR was used to detect mRNA of Bax and Bcl-2 genes, and Western blot was used to detect the protein of Bax and Bcl-2. Furthermore, cell apoptosis rate was estimated by flow cytometry. Results Compared with the control group, the expressions of Bcl-2 mRNA and protein decreased ( P < 0. 05 ), and the expressions of Bax mRNA and protein in different DEHP groups increased ( P < 0. 05 )in a concentration dependent manner respectively. Meanwhile, the cell apoptosis increased( P < 0. 05 ) in a concentration dependent manner. Conclusion The DEHP could induce apoptosis in rat Leydig cells by up-regulating the Bax gene expression and down-regulating the Bcl-2 gene expression, which results in Leyding cell dysfunction.%目的 通过对原代培养的大鼠睾丸间质细胞进行邻苯二甲酸二乙基己酯(di-2-ethylhexyl phthalate,DEHP)染毒,探讨DEHP体外对大鼠睾丸间质细胞Bax和Bcl-2基因表达及凋亡的影响.方法 体外分离和原代培养未成熟大鼠睾丸间质细胞,用不同浓度的DEHP(10、50、100nmol/L)染毒24h.荧光定量PCR法检测间质细胞Bax和Bcl-2基因mRNA表达,Weastern blot检测间质细胞Bax和Bcl-2蛋白表达,流式细胞术检测间质细胞凋亡率.结果 与对照组相比,DEHP各组睾丸间质细胞Bcl-2基因mRNA和蛋白表达下降(P＜0.05),Bax基因mRNA和蛋白表达增加(P＜0.05),细胞凋亡率增加(P＜0.05),呈浓度依赖关系.结论 DEHP可通过抑制Bcl-2基因表达和上调Bax基因表达诱导大鼠睾丸间质细胞凋亡,进而影响间质细胞功能.