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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、Claudin-11mRNA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和Claudin-11 mRNA表达。结果 1与空白对照组比较,各染锰组支持细胞数量及VM阳性细胞率、Occludins mRNA和Claudin-11 mRNA表达均显著降低。2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,支持细胞数量及VM阳性细胞率、Occludins mRNA和Claudin-11 mRNA表达均显著降低。3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,破坏血睾屏障,导致生精微环境改变,产生生殖毒性效应。
Objective To investigate the ultrastructural changes of testis induced by manganese and the expression of vimentin (VM) and tight junctions of Occludins and Claudin-11 mRNA in rats, and to explore the mechanism of the destruction of cytoskeleton proteins and claudin 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 Occludins and Claudin-11 mRNA in the blood-testis barrier was detected by real-time quantitative PCR. Results 1 Compared with the blank control group, the number of supporting cells and the rate of VM-positive cells in each manganese-treated group were significantly lower than those in the blank control group. The expressions of both Occludins mRNA and Claudin-11 mRNA were significantly decreased. The results showed that the number of supporting cells and the rate of VM positive cells in the high dose group and the low dose group were significantly lower than those in the low dose group at 6 weeks and 4 weeks, and the rates of Occludins mRNA and Claudin-11 mRNA were significantly decreased. 3 The number of testicular sertoli cells in each group was positively correlated with the rate of VM positive cells and the expressions of Occludins mRNA and Claudin-11 mRNA. Conclusion Manganese can inhibit the expression of skeletal proteins and tight junction-related proteins in rat testicular sertoli cells, destroy the blood-testis barrier, lead to the changes of sperm microenvironment and produce reproductive toxicity.