抗肿瘤药物环磷酰胺对大鼠睾丸和精原干细胞相关因子表达的影响分析

Effect of anti-tumor drugs cyclophosphamide on the expression of related factors in testis and sperm of rats

目的 探讨抗肿瘤药物环磷酰胺（cyclophosphamide,CTX）对雄性大鼠睾丸、精原干细胞及相关因子八聚体结核转录因子4（OCT-4）、胶质细胞源神经营养因子（glial cell line derived neurotrophic factor,GDNF）表达的影响.方法 选取16只8周龄Wister雄性大鼠采用随机数字表法分为CTX组和对照组各8只,CTX组采取连续3d腹腔注射CTX（50 mg/kg）,对照组采取等量生理盐水,对比两组大鼠给药后1、2、4周的精子细胞凋亡数及S期细胞所占比例的变化;对比两组大鼠给药4周后的睾丸、附睾重量、生精小管直径差异;采用免疫组化染色检测两组睾丸OCT-4、GDNF蛋白的表达情况.结果 CTX组大鼠在给药后1、2、4周的生精细胞凋亡数均显著的高于同期的对照组（P＜0.05）;CTX组大鼠在给药后1、2、4周的生精细胞S期所占细胞比例均显著的低于同期的对照组（P＜0.05）;CTX组大鼠在给药4周的睾丸、附睾重量显著地低于同期的对照组（P＜0.05）;两组大鼠的生精小管直径差异无统计学意义（P＞0.05）;CTX组大鼠睾丸组织中GDNF蛋白阳性表达显著地低于对照组大鼠（P＜0.05）;两组大鼠的OCT-4蛋白表达差异无统计学意义（P＞0.05）.结论 CTX具有诱导精原细胞凋亡的作用,干扰精原细胞增殖分化功能,可能与下调睾丸组织中GDNF有关.

Objectives To investigate the effect of cyclophosphamide （CTX） on the expression of the octamer transcription factor-4 （OCT-4） and glial cell line derived neurotrophic factor （GD-NF） in the testis,the primary stem cell and the related factor in male rats.Methods Sixteen 8-week-old male Wister rats were randomly divided into CTX group and control group（with 8 animals in each group）,CTX group received intraperitoneal injection of continuous 3 d CTX （50 mg/kg）,the control group received normal saline.The number of apoptotic sperm cells and the proportion of cells in s phase were compared between the two groups after 1,2 and 4 weeks of medication.Testis and epididymis weight,seminiferous tubule diameter were compared between the two groups after 4 weeks.OCT -4,GDNF protein expression was detected by immunohistochemical staining.Results Apoptotic spermatogenic cells in CTX group after 1,2 and 4 weeks of medication were significantly higher than the same period in the control group （P 〈 0.05）.Germ cell percentage of s phase cells after 1,2 and 4 weeks of medication in the CTX group was significantly lower than the same period in the control group （P 〈 0.05）.Testis and epididymis weight in the CTX group after 4 weeks were significantly lower than the same period the control group （P 〈 0.05）.There were no statistically significant differences in diameter of seminiferous tubules between the two groups （P 〉 0.05）.GDNF protein expression of testes in CTX group was significantly lower than the control group （P 〈 0.05）.There was no statistically significant difference in OCT-4 protein expression between both groups （P 〉 0.05）.Conclusions CTX spermatogonia induces apoptosis,interferes with spermatogonial cell proliferation and differentiation,may be related with down-regulating GDNF in testicular tissue.