摘要
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凋亡通路Caspase-3和Caspase-9表达以及细胞凋亡的影响。方法:体外分离和原代培养大鼠睾丸间质细胞,用不同浓度的DEHP(10、50、100 nmol/L)染毒24 h。荧光定量PCR法检测间质细胞Caspase-3和Caspase-9 mRNA表达,Western印迹检测间质细胞Caspase-3和Caspase-9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Caspase-3 mRNA(1.69±0.38 vs3.82±0.39、6.91±0.40、15.47±0.40)和蛋白(0.18±0.09 vs 0.32±0.10、0.61±0.08、0.89±0.09)表达显著增加(P均<0.05),Caspase-9 mRNA(2.24±0.41 vs 5.16±0.43、9.61±0.45、19.22±0.43)和蛋白(0.26±0.07 vs0.40±0.08、0.68±0.09、0.96±0.08)表达也显著增加(P均<0.05),细胞凋亡率(4.36±1.11 vs 7.52±1.09、12.72±1.10、24.59±1.11)也显著增加(P均<0.05),呈浓度依赖关系。结论:DEHP可通过激活细胞凋亡Caspase通路而诱导大鼠睾丸间质细胞凋亡,进而影响间质细胞功能。
Objective： To investigate the effects of di-（2-ethylhexyl） phthalate （DEHP） on the expressions of Caspase-3 and Caspase-9 genes in rat Leydig cells and the apoptosis of the cells in vitro. Methods ： Leydig cells were isolated from male SD rats, primarily cultured and treated with DEHP at a low （10 nmol/L）, a medium （50 nmol/L） and a high dose （100 nmol/L） for 24 hours. Then the mRNA expressions of Caspase-3 and Caspase-9 genes in the Leydig cells were detected by real time PCR, their protein expressions determined by Western blot, and the apoptosis of the Leydig cells measured by flow eytometry. Results ： Compared with the DMSO control group, the low-, medium- and high-dose DEHP groups showed significantly upregulated expressions of Caspase-3 mRNA （ 1.69 ± 0.38 vs 3.82 ± 0.39, 6.91 ± 0.40 and 15.47 ± 0.40, P 〈 0.05 ）, Caspase-3 protein （0. 18 ± 0.0.09 vs 0. 32 ± 0.10, 0.61 ± 0.08 and 0.89 ± 0.09, P 〈 0.05）, Caspase-9 mRNA （2.24 ± 0.41 vs 5.16 ± 0.43,9.61 ± 0.45 and 19.22 ± 0.43, P 〈 0.05 ） and Caspase-9 protein （ 0.26 ± 0.07 vs 0. 40 ± 0.08, 0.68 ± 0.09 and 0.96 ± 0.08, P 〈 0.05 ）, as well as increased apoptosis rate of Leydig cells （4.36 ± 1.11 vs 7.52 ±1.09, 12.72 ± 1.10 and 24.59 ± 1.11, P 〈0.05）, all in a dose-dependent manner. Conclusion ： DEHP can induce the apoptosis of rat Leydig cells by activating the apoptosis Caspase pathway, and consequently affect the function of Leydig cells.
出处
《中华男科学杂志》
CAS
CSCD
2013年第8期683-688,共6页
National Journal of Andrology
基金
天津市职业与环境危害生物标志物重点实验室开放基金(WZK200905)~~
