GDNF干预对化疗致精原干细胞耗竭小鼠生精细胞恢复效果的研究

THE STUDY OF THE EFFECT OF GDNF INTERVENTION ON THE RECOVERY OF SPERMATOGENIC CELLS IN THE CHEMOTHERAPY INDUCED SPERMATOGONIAL STEM CELL DEPLETED MICE

目的:研究外源胶质细胞源性神经营养因子(glialcell line-derived neurotrphic factor,GDNF)对化疗致不育小鼠生精细胞的恢复效果,明确GDNF促进在体精原干细胞再生的效果。方法:4周龄雄性C57小鼠腹腔注射白消安(30mg/kg),注射后4周按60μg/kg体重剂量腹腔注射GDNF进行干预,每天一次连续注射7d。在干预后第4周收集睾丸,检测睾丸重量、通过HE染色检测生精小管中生殖细胞恢复情况;通过RT-PCR检测GDNF和SCF基因表达评价GDNF注射对小鼠精原干细胞微环境的影响。结果:在GDNF干预后第4周,睾丸重量明显得到恢复,生精小管中上皮细胞数量明显得到恢复,精原干细胞微环境中再生因子GDNF和分化因子SCF表达水平稳定。结论:GDNF干预可以促进化疗致精原干细胞耗竭小鼠睾丸中生精细胞数量明显恢复,且维持了精原干细胞微环境中再生分化调控的平衡关系。

Objective：To study the effect of glial cell line - derived neurotrphic factor （GDNF） on the recovery of spermatogenic cells in the chemotherapy induced infertile mice, and clarify the effectiveness of exogenous GDNF on promoting regeneration of spermat- ogonial stem cells in vivo. ＇Methods： Four - week - old male C57 mice were injected with busulfan via abdomen （30 mg per kilogram of body weight）. After four weeks, the mice received the intraperitoneal injection of GDNF, 60 μg per kilogram of body weight and once a day lasting for 7 days In the fourth week after intervention, the testis of the mice were collected and weighted also the recovery of germ cells in seminiferous tubules was detected via HE staining. The expression of GDNF gene and SCF gene were detected by RT -PCR, and the effect of GDNF intervention on the mi of spermatogonial stem cells was estimated. Results： In the fourth week following GDNF intervention, the weight of the testis was recovered, the amount of epithelial ceils in seminiferous tubules was recovered obviously, and the expression of regeneration factor GDNF and differentiation factor SCF are stable in the niche of sper- matogonial stem cells. Conclusion： The GDNF intervention could promote the significant recovery of spermatogenic cell number in the testis of mice , whose spermatogonial stem cell depletion was induced by chemotherapy,and the balance between regeneration and differ- entiation in the niche of spermatogonial stem cells was maintained as well.