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目的探讨锰对大鼠生精细胞Caspase-3 mRNA调控及支持细胞波形蛋白(vimentin)表达的影响。方法将正常雄性SD大鼠48只,随机分为1个对照组和2个染锰组,给予各组大鼠腹腔注射生理盐水和15mg/kg、30mg/kg氯化锰。分别染锰4周和6周,随机处死各组大鼠8只。应用末端脱氧核糖核酸转移酶介导的dUTP缺口末端标记技术(TUNEL)、原位杂交法和免疫组织化学链酶亲和素-生物素-过氧化物酶复合物(SABC)法进行检测研究。结果 1.与空白对照组比较,15mg/kg、30mg/kg染锰组生精细胞凋亡指数(AI)均升高(P<0.05,P<0.01),Caspase-3 mRNA阳性细胞率均显著升高(P<0.01),支持细胞波形蛋白(vimentin)阳性细胞率均显著降低(P<0.01)。2.染锰剂量相同,染锰6周组与染锰4周组比较,生精细胞AI与Caspase-3 mRNA阳性细胞率均显著升高(P<0.01),支持细胞vimentin阳性细胞率均显著降低(P<0.01)。3.染锰时间相同,30mg/kgMnCl2组与15mg/kg MnCl2组比较,生精细胞AI与Caspase-3 mRNA阳性细胞率均显著升高(P<0.01),支持细胞vimentin阳性细胞率均显著降低(P<0.01)。4.各组大鼠生精细胞AI和Caspase-3 mRNA阳性细胞率呈正相关(r=0.842,P<0.01),与支持细胞vimentin阳性细胞率呈负相关(r=-0.859,P<0.01),各组大鼠生精细胞Caspase-3 mRNA阳性细胞率和支持细胞vimentin阳性细胞率呈负相关(r=-0.975,P<0.01)。结论染锰大鼠生精细胞Caspase-3 mRNA表达增加导致生精细胞凋亡增加;支持细胞vimentin表达下降;这可能是锰生殖毒性的重要分子机制之一。
Objective To investigate the effects of manganese on the regulation of Caspase-3 mRNA and the expression of vimentin in rat spermatogenic cells. Methods Forty-eight normal male SD rats were randomly divided into 1 control group and 2 manganese-exposed groups. Rats in each group were injected intraperitoneally with normal saline and 15 mg / kg and 30 mg / kg of manganese chloride. Manganese were dyed for 4 weeks and 6 weeks respectively, and 8 rats in each group were randomly sacrificed. End-to-end DNT-mediated dUTP nick end labeling (TUNEL), in situ hybridization and immunohistochemical streptavidin-biotin-peroxidase complex (SABC) assays were performed. Compared with the blank control group, the apoptosis index (AI) of spermatogenic cells in 15mg / kg, 30mg / kg Mn-Mn group increased (P <0.05, P <0.01) and the positive rate of Caspase- (P <0.01), and the percentage of vimentin positive cells decreased significantly (P <0.01). Compared with the 4-week group, the AI rates of AI and Caspase-3 mRNA in sperm cells were significantly increased (P <0.01), and the positive rate of vimentin in supportive cells was significantly higher Decreased (P <0.01). The same time of manganese dying, the positive cells of AI and Caspase-3 mRNA of spermatogenic cells in 30mg / kg MnCl2 group and 15mg / kg MnCl2 group were significantly increased (P <0.01), and the percentage of vimentin positive cells in supporting cells was significantly lower (P <0.01). The positive rates of AI and Caspase-3 mRNA of spermatogenic cells were positively correlated (r = 0.842, P <0.01), and negatively correlated with the rate of vimentin positive cells (r = -0.859, P <0.01) The positive rate of Caspase-3 mRNA in spermatogenic cells was negatively correlated with the rate of vimentin positive cells (r = -0.975, P <0.01). Conclusion The increased expression of Caspase-3 mRNA in spermatogenic cells of manganese-exposed rats resulted in increased apoptosis of spermatogenic cells and decreased expression of vimentin in supportive cells. This may be one of the important molecular mechanisms of manganese reproductive toxicity.