三氯异氰尿酸对SD大鼠睾丸生精细胞凋亡相关蛋白表达的影响

Effects of trichloroiminocyanuric acid on apoptosis-associated proteins of spermatogenic cells in SD rats

目的观察凋亡相关蛋白Bcl-2/Bax、Fas/FasL、Caspase-3在三氯异氰尿酸(TCCA)染毒大鼠睾丸中的表达,探讨其在TCCA诱导生精细胞的凋亡中发挥的作用。方法 30只SPF级别雄性SD大鼠分为3组,每组10只。按0、275和550 mg/kg剂量隔日经口灌胃10周后,处死大鼠,免疫组织化学法检测细胞凋亡相关蛋白Bcl-2/Bax、Fas/FasL、Caspase-3的表达,实时荧光定量PCR法检测Bcl-2 mRNA和Fas mRNA的表达水平。结果 TCCA染毒组Bcl-2、Bax蛋白表达降低,Bcl-2/Bax的比值降低,Fas、FasL蛋白表达增加,Caspase-3蛋白表达增加,同时睾丸中Bcl-2 mRNA和Fas mRNA表达水平升高。结论 Bcl-2/Bax、Fas/FasL介导的2种凋亡途径在TCCA诱导的雄性生殖毒性中均发挥一定作用。

Objective To evaluate the expression of cell apoptosis related factors （Bcl-2/Bax,Fas/FasL and Caspase-3 protein） in testis of rats exposed to trichloroiminocyanuric acid （ TCCA ） and investigate their effects on the spermatogenic cells apoptosis induced by TCCA. Methods 30 male rats were divided into three groups and were given a gavage of TCCA at the dosage of 0,275,550mg/kg for 10 weeks （each other day） respectively. Immunohistochemistry was used to observe the change of these proteins and real-time fluorescence quantitative PCR was used to observe the gene expression level. Results The expression of Fas/FasL increased whereas that of Bcl-2/Bax decreased with the radio of Bcl-2/Bax. Caspase-3 protein was activated while the expression levels of Fas mRNA and Bcl-2 mRNA were significantly higher in testis of 275mg/kg group. Conclusion The two apoptosis pathways mediated by Fas/FasL and Bcl-2/Bax possibly participate in the process of the male reproductive toxicity induced by TCCA.