孔雀石绿对Fas/Fas-L诱导的小鼠睾丸生精细胞凋亡的影响

目的:探讨孔雀石绿对小鼠睾丸生精细胞由Fas/Fas-L所诱导凋亡的影响。方法:32只雄性成年BALB/c小鼠随机均分为4组,按照孔雀石绿浓度设为:对照组(A组,0mg/L)、低剂量组(B组,100mg/L)、中剂量组(C组,400mg/L)、高剂量组(D组,800mg/L)。实验组连续灌胃给药30d,对照组以等量生理盐水代替孔雀石绿。HE染色后观察睾丸组织结构的变化,免疫组化法检测Fas及Fas-L蛋白的表达及改变。结果:B组显示与对照组相似的组织学结构。C组和D组的生精上皮细胞层数减少,少数管腔内可见脱落生精细胞,病理变化随剂量增大而加重。Fas和Fas-L在间质细胞和精子的表达呈强阳性,Fas集中于精子细胞核及周围残余胞浆,管腔中的絮状精子尾明显见Fas-L表达。Fas和Fas-L在支持细胞表达较弱,在A、B组表达位于靠近基底膜处的胞浆,在C、D组还表达于伸向管腔的胞浆。Fas在C、D组精子及支持细胞的表达均比对照组显著增强,Fas-L在3个实验组支持细胞的表达均显著强于对照组(P<0.05),在精子表面的表达则无明显差异。结论:孔雀石绿可以导致小鼠睾丸生精上皮损伤和支持细胞的Fas/Fas-L表达上调,表明孔雀石绿可能通过Fas/Fas-L通路使生精细胞凋亡增加。

Effects of malachite green on apoptosis of spermatogenic cells induced by Fas/Fas-L in the testis of mouse

Objectives: To investigate effects of malachite green (MG) on apoptosis of spermatogenic cells induced by Fas/Fas-L, and evaluate the injuries of testis caused by MG. Methods: Thirty-two male BALB/c mice were randomly divided into 4 groups: A, B, C and D. MG solved in normal saline was given at the dose of 0, 100, 400, or 800 ppm in each group for 30 days by intragastric administration consecutively. Changes of testicular pathology were observed by light microscopy, and the expression of both Fas and Fas-L was detected by using immunohistochemistry. Results: Obvious changes of testicular pathology appeared in groups C and D, but not in group B. Both Fas and Fas-L expressed in sperm and Leydig cells were strong while they were weak in Sertoli cells. Fas concentrated in sperm nuclei and the slough near sperm, and Fas-L located on sperm tails. Fas expressed in Sertoli cells and sperm increased significantly in groups C and D. The positive expression of Fas-L in Sertoli cells was stronger in all treated groups than that in the control (P<0.05). Conclusion: MG would have deleterious effects on testicular structures of mouse, and could enhance apoptosis of spermatogenic cells by increasing Fas/Fas-L, which led to decrease spermatogenesis.