染锰大鼠生精细胞凋亡中caspase-3 mRNA、凋亡蛋白酶活化因子-1及多聚ADP核糖聚合酶的表达变化  

Expression changes of caspase-3 mRNA,apoptosis protease activating factor-1 and poly ADP-ribose polymerase in apoptosis of spermatogenic cells of rats exposed to manganese

作　　者：郭海 宋爽 王国秀[2] 才秀莲[2] 

机构地区：[1]河南医学高等专科学校组织学与胚胎学教研室,郑州451191 [2]遵义医学院组织学与胚胎学教研室,遵义563003

出　　处：《解剖学杂志》2017年第3期254-257,F0004,共5页Chinese Journal of Anatomy

基　　金：遵义市科技项目基金(200707);遵义市红花岗区科技项目基金(200622)

摘　　要：目的:研究染锰诱导的大鼠生精细胞凋亡过程中,天冬氨酸特异性半胱氨酸酶-3(caspase-3)mRNA和凋亡蛋白酶活化因子-1(Apaf-1)、多聚ADP核糖聚合酶(PARP)的表达及其关系,探讨它们在生精细胞凋亡过程中的作用。方法:雄性SD大鼠,设立空白对照组、低剂量(15 mg/kg MnCl_2)和高剂量(30 mg/kg MnCl_2)组。实验组分别染锰4周和6周,空白对照组给予等容生理盐水,给药途径均为腹腔注射,TUNEL法检测生精细胞凋亡,原位杂交法和免疫组织化学法检测生精细胞caspase-3 mRNA、Apaf-1和PARP的表达。结果:与空白对照组比较,各染锰组生精细胞凋亡指数(AI)和caspase-3mRNA阳性细胞率均显著升高,Apaf-1和PARP阳性细胞率均显著降低。染锰剂量相同,6周与4周组比较,以及染锰时间相同,高剂量组与低剂量组比较,生精细胞AI和caspase-3 mRNA阳性细胞率均显著升高,Apaf-1和PARP阳性细胞率均显著降低。各组大鼠生精细胞AI与caspase-3 mRNA阳性细胞率呈正相关,与Apaf-1和PARP阳性细胞率呈负相关。结论:锰可影响大鼠生精细胞caspase-3 mRNA表达,促进Apaf-1和PARP分解,导致生精细胞凋亡,产生生殖毒性效应。Objective： To determine the effect of caspase-3 mRNA and apoptosis protease activating factor-1 （Apaf-1）, poly ADP-ribose polymerase （PARP） in apoptosis of spermatogenic cells of rats exposed to manganese. Methods： SD male rats were randomly divided into one control group and two manganese chloride （MnCl2） （15 mg/kg, 30 mg/kg） groups respectively. After being exposed to manganese for 4 and 6 weeks, the apoptosis of spermatogenic cells was examined by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling （TUNEL） technique. The expression of caspase-3 mRNA in spermatogenic cells was investigated by in situ hybridization, and the expression of Apaf-1 and PARP was detected by immunohistochemistry. Results： Compared with the control group, AI and caspase-3 mRNA-positive-cell rates were L increased but the Apaf-1 and PARP-positive-celI rates were decreased. Among the same dose and same time manganese exposure groups, AI and caspase-3 mRNA-positive-cell rates were increased but the Apaf-1 and PARP-positive-cell rates were decreased in the 15 mg/kg MnC12 group compared to the 30 mg/kg MnCl2 group. There existed a positive correlation between AI and the caspase-3 mRNA-positive-cell rate and a negative correlation between AI and the Apaf-l-positive-cell rate and PARP-positive-cell rate. Conclusion： Manganese could upregulate the ,expression of caspase-3 mRNA, and decompose Apaf-1- positive-cell rate and PARP-positive-cell rate, which might be one of the important molecular mechanisms of reproductive toxicity of manganese.

关 键 词：锰 生精细胞 凋亡 天冬氨酸特异性半胱氨酸酶-3 MRNA 凋亡蛋白酶活化因子-1 多聚ADP核糖聚合酶 

分 类 号：R114[医药卫生—卫生毒理学]