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目的研究染锰诱导的大鼠睾丸超微结构改变及支持细胞vimentin(VM)和紧密连接Occludins mRNA和Claudin-11 mRNA表达,探讨锰对支持细胞骨架蛋白和紧密连接蛋白的破坏机制。方法雄性SD大鼠随机分为空白对照组,低剂量(15 mg/kg MnCl_2)和高剂量(30 mg/kg MnCl_2)组,8只/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,电镜观察睾丸支持细胞及血睾屏障超微结构,免疫组织化学(SABC)法检测支持细胞VM表达,实时定量PCR反应检测血睾屏障紧密连接Occludins,Claudin-11 mRNA表达。结果随着染锰时间延长和剂量增加,各组支持细胞数量及VM阳性细胞率、Occludins mRNA和Claudin-11 mRNA表达水平均显著降低。各组大鼠睾丸支持细胞数量与VM阳性细胞率及Occludins mRNA和Claudin-11 mRNA表达水平均成正相关。结论锰可抑制大鼠睾丸支持细胞骨架蛋白及紧密连接相关蛋白表达,产生生殖毒性效应。
Objective To investigate the ultrastructural changes of testis induced by manganese in rats and the expression of vimentin (VM) and tight junction Occludins mRNA and Claudin-11 mRNA in rat testis, and to explore the mechanism of the destruction of cytoskeleton and claudin binding proteins. Methods Male Sprague-Dawley rats were randomly divided into blank control group, low dose (15 mg / kg MnCl 2) and high dose (30 mg / kg MnCl 2) group, 8 rats / group. The experimental group were drank manganese for 4 weeks and 6 weeks, respectively, and the blank control group was given isocratical saline. The route of administration was intraperitoneal injection. The ultrastructure of sertoli cells and blood-testis barrier was observed by electron microscopy and immunohistochemistry (SABC) The expression of Claudin-11 mRNA and Occludins were detected by real-time PCR. Results With the prolongation of manganese dose and dose, the number of supportive cells and the percentage of VM positive cells in each group were significantly decreased. The number of testicular sertoli cells in each group was positively correlated with the rate of VM positive cells and the expression level of Occludins mRNA and Claudin-11 mRNA. Conclusion Manganese can inhibit the expression of cytoskeleton proteins and tight junction-related proteins in rat testis and produce reproductive toxicity.