目的:观察胚胎期氟他胺(Flu)对SD大鼠睾丸生殖细胞发育的影响,建立研究隐睾生殖母细胞(Go)发育缺陷机制及治疗的模型。方法:36只SD孕鼠随机分为Flu组(n=20)、玉米油组(n=8)和空白对照组(n=8)3组,Flu组和玉米油组于妊娠12~21 d(GD12-21)分别给予Flu 25 mg/(kg.d)和1 ml/(kg.d)皮下注射,在出生后1 d(PD1)、PD10、PD20、PD80收集标本,观察睾丸形态学、组织学的差异,免疫组化及RT-PCR检测神经母细胞粘附分子(NCAM)表达。结果:Flu诱导隐睾发生率为43.9%(29/66)。PD20、PD80隐睾睾丸质量、脏器系数较对照组有显著性差异;PD10 Flu诱导睾丸组织中可见Go迁移障碍,滞留于管腔中央,Flu诱导PD20、PD80隐睾组织中仍可见迁移障碍的Go位于管腔中央;免疫组化检测结果显示迁移障碍的Go胞膜阳性表达NCAM,且PD10、PD20睾丸RT-PCR检测结果显示Flu诱导隐睾组织中NCAM的mRNA表达高于对照组。结论:Flu成功诱导大鼠隐睾Go残留模型,可以用于研究隐睾生殖母细胞发育缺陷的机制及治疗。
Objective:To observe the effects of flutamide(Flu) on the development of testicular germocytes in SD rats,and to establish a rat model for further researches on the maldevelopment of cryptorchidism gonocytes(Go).Methods:Pregnant SD rats were subcutaneously injected with Flu from gestational day(GD) 12 to 21 to establish a model of cryptorchidism.The testes of the newborns were harvested on postnatal day(PD) 1,10,20 and 80 for observation of their morphological and histological changes by HE staining and detection of the expression of neural cell adhesion molecules(NCAM) by immunohistochemistry and RT-PCR.Results:Flu induced 43.9%(29/66) of cryptorchidism in the exposed rats.Significant differences were found in the testicular weight and organ coefficient between the Flu and the control groups on PD 20 and 80.Gos remained in the center of seminiferous tubules of the Flu-induced testis on PD 10,and in the center of seminiferous tubules in the cryptorchids' testicular tissues on PD 20 and 80.Immunohistochemistry showed the expression of NCAM on the membrane of the remaining Gos,and RT-PCR revealed significantly up-regulated expression of NCAM mRNA in the Flu-induced testes on PD 10 and 20 as compared with the controls(P0.05).Conclusion:A rat model of Flu-induced cryptorchidism with remaining Gos was successfully established,which could be used to study the mechanism and management of the maldevelopment of cryptorchidism gonocytes.