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目的研究锰对大鼠生精细胞中半胱氨酰天冬氨酸酶-3(caspase-3)和细胞色素C(cytochrome-c)表达的影响。方法将健康雄性清洁级SD大鼠48只随机分为高(30mg/kg)、低剂量(15mg/kg)MnCl2染毒组和空白对照组(生理盐水),每组16只。采用腹腔注射方式染毒,每天1次,每周5天,连续染毒6周。分别于染毒第4、6周末称重,处死大鼠,分离睾丸,采用免疫组化法检测睾丸生精细胞中caspase-3与cytochrome-c的阳性细胞率。并进行病理学观察。结果与空白对照组比较,高、低剂量MnCl2染毒组大鼠生精细胞中caspase-3阳性细胞率均升高,cytochrome-c阳性细胞率均降低,差异有统计学意义(P<0.01);且随着锰染毒剂量的升高,大鼠生精细胞中caspase-3阳性细胞率呈上升趋势,cytochrome-c阳性细胞率呈下降趋势。与染锰第4周比较,高、低剂量MnCl2染毒组第6周大鼠生精细胞中caspase-3阳性细胞率均升高,cytochrome-c阳性细胞率均降低,差异有统计学意义(P<0.01)。直线相关分析发现,各组大鼠生精细胞caspase-3阳性细胞率与cytochrome-c阳性细胞率呈显著负相关(r=-0.876,P<0.01)。结论染锰可通过线粒体途径促进大鼠生精细胞caspase-3表达,而降低cytochrome-c表达,且存在一定的剂量-效应和时间-效应关系。这可能是导致大鼠生精障碍的主要机制之一。
Objective To study the effects of manganese on the expression of caspase-3 and cytochrome-c in rat spermatogenic cells. Methods Forty-eight healthy male SD rats were randomly divided into high (30mg / kg), low dose (15mg / kg) MnCl2 groups and blank control group (normal saline), 16 rats in each group. Using intraperitoneal injection of exposure, once daily, 5 days a week, continuous exposure to 6 weeks. The rats were sacrificed at the end of the 4th and 6th week respectively, and the testis was isolated. The positive rate of caspase-3 and cytochrome-c in testicular spermatogenic cells was detected by immunohistochemistry. And pathological observation. Results Compared with the blank control group, the rates of caspase-3 positive cells and cytochrome-c positive cells of spermatogenic cells in high and low dose MnCl2 groups were significantly decreased (P <0.01) ; And with the increase of manganese exposure dose, the rate of caspase-3 positive cells in rat spermatogenic cells showed an upward trend and the rate of cytochrome-c positive cells showed a decreasing trend. Compared with the 4th week of manganese doping, the rates of caspase-3 positive cells and the cytochrome-c positive cells in spermatogenic cells of rats in the high and low dose MnCl2 groups were all significantly decreased at the 6th week P <0.01). Linear correlation analysis showed that there was a significant negative correlation between the rate of caspase-3 positive cells and the percentage of cytochrome-c positive cells (r = -0.876, P <0.01). Conclusion Manganese can promote the expression of caspase-3 in rat spermatogenic cells through mitochondrial pathway and decrease the expression of cytochrome-c, and there is a certain dose-effect and time-effect relationship. This may be one of the main mechanisms that lead to spermatogenesis in rats.