环磷酰胺干扰精原干细胞功能的初步研究

目的:初步探讨环磷酰胺对精原干细胞分化功能的干扰作用。方法:根据W istar大鼠生精细胞发育不同阶段分为1、3、9周龄组,每组24只,随机均分为实验组和对照组,实验组腹腔内注射环磷酰胺100 mg/(kg.体重),对照组注射等量生理盐水,24 h后用原位缺口末端标记法检测生精细胞凋亡。另取仅有精原干细胞的1周龄W istar雄性大鼠60只,随机均分为实验组和对照组(给药方法同上),在给药后24 h、3周、9周分别用免疫组化法检测c-K it蛋白,流式细胞仪分析细胞周期。结果:环磷酰胺干预后,1周龄实验组与对照组大鼠生精细胞凋亡差异无显著性(P>0.05),其余各周龄组实验组生精细胞凋亡均显著增高(P<0.01)。1周龄实验组大鼠睾丸在环磷酰胺处理后不同时间点的细胞周期S期细胞比例、精原细胞c-K it蛋白表达水平均明显低于对照组(P<0.01)。结论:环磷酰胺诱导精原干细胞凋亡不明显,可能更重要的是干扰精原干细胞的增殖分化功能。

Effect of Cyclophosphamide on Spermatogonial Stem Cells

Objective: To perform a preliminary study on the dysfunction of spermatogonial stem cells(SSCs) induced by cyclophosphamide.Methods: According to development stage of spermatogenesis in Wistar rat,72 rats were divided into three groups,1,3 and 9 weeks groups, respectively.Then 24 rats per group were randomly divided into experimental and control groups,and 100 mg/kg cyclophosphamide was injected by intraperitoneal injection in experimental groups,with the same volume of normal saline in the control.Apoptosis of spermatogenic cells were detected after 24 hours by TUNEL. Then 60 one-week rats,whose germ cells were only SSCs,were randomly divided into experimental and control group.We detected c-Kit by immunohistochemistry and cell cycle by flow cytrmetry at 24 hours,3 and 9 weeks.Results: There was no significant difference in apoptosis of germ cells in 1 week between experimental group and control group(P>(0.05));however,there were significant differences among other groups.The ratio of S stage and c-Kit expression of spermatogonial cells were decreased in one-week rats at 24 hours, 3 and 9 weeks in experimental group compared with the control,respectively(P<(0.01)).Conclusion: Cyclophosphamide does not significantly induce SSCs apoptosis.It may be more important to interfere the proliferation and differentiation of SSCs.