锰诱导大鼠生精细胞凋亡过程中半胱氨酸酶-3mRNA与增殖细胞核抗原表达变化  

Effect of manganese on the expression of caspase-3 mRNA and PCNA in rat spermatogenic cells apoptosis

作　　者：郭海 才秀莲[2] 王国秀[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》2015年第1期15-17,103,F0003,共5页Chinese Journal of Anatomy

基　　金：遵义市科技项目基金[遵市科合社字(200707号)];遵义市红花岗区科技项目基金[遵红科合社字(200622号)]

摘　　要：目的：研究氯化锰对生精细胞凋亡过程半胱氨酸酶3 （caspase-3） mRNA和增殖细胞核抗原（PCNA）表达的影响,探讨两者在生精细胞凋亡过程中的作用.方法：雄性SD大鼠随机分为空白对照组、低剂量（15 mg/kg MnC12）和高剂量（30 mg/kg MnCl2）组.MnCl2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法检测睾丸生精细胞凋亡,原位杂交和免疫组织化学（SABC）检测生精细胞caspase-3 mRNA和PCNA和表达.结果：与空白对照组比较,各染锰组生精细胞凋亡指数（AI）均升高,caspase-3 mRNA阳性细胞率均显著升高,染锰4周PCNA阳性细胞率显著降低,染锰6周反而升高.染锰剂量相同,6周与4周组比较,生精细胞AI与caspase-3 mRNA阳性细胞率、PCNA阳性细胞率均显著升高.染锰时间相同,高剂量组与低剂量组比较,生精细胞AI与caspase-3 mRNA阳性细胞率均显著升高,PCNA阳性细胞率显著降低.各组大鼠生精细胞AI与caspase-3 mRNA阳性细胞率呈正相关,与PCNA阳性细胞率呈负相关,而caspase-3 mRNA阳性细胞率与PCNA阳性细胞率呈负相关.结论：锰可诱导大鼠生精细胞caspase-3mRNA表达,促使生精细胞凋亡并且抑制细胞增殖,产生生殖毒性效应.Objective： To determine the effect of manganese on the expression of caspase-3 mRNA and PCNA in apoptosis of rat spermatogenic cells. Methods： Male rats were randomly divided into one control group and two manganese chloride （MnClz） （15 mg/kg, 30 mg/kg） groups. After exposure 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 PCNA was detected by immunohistochemistry （SABC）. Results： Compared with the control groups, AI was increased, the caspase- 3 mRNA-positive-cell rate was increased significantly, and the PCNA-positive-cell rate was decreased in 4-week group but increased in 6-week group significantly. Among same dose manganese exposure groups, AI, caspase-3 mRNAi-positive-cell rate and PCNA-positive-cell rate were increased more significantly in the 6-week group than that in the 4-week group. Among manganese exposure groups of the same time, AI and caspase-3 mRNA-positive-cell rate in 30 mg/kg manganese exposure groups were increased more significantly than that in 15 mg/kg manganese exposure groups, but the PCNA-positivesfell rate was decreased more significantly in 30 mg/kg manganese exposure groups than that in 15 mg/kg manganese exposure groups. There existed a positive correlation between AI and the caspase-3 mRNA-positive-cell rate, a negative correlation between AI and PCNA-positivecell rate, and a negative correlation between the caspase-3 mRNA-positive-cell rate and the PCNA- positive-cell rate. Conclusion： After being exposed to manganese, the increased expression of caspase-3 mRNA in spermatogenic cells led to increased apoptosis and decreased proliferation of spermatogenic cells in rats, which might be one of the most important molecular mechanisms of reproductive toxicity of manganese.

关 键 词：锰 生精细胞 凋亡 CASPASE-3 MRNA 增殖细胞核抗原 大鼠 

分 类 号：R114[医药卫生—卫生毒理学]