大鼠隐睾模型的建立及胚胎干细胞原位注射的治疗研究  

Establishment of rat cryptorchidism model and orthotopic injection of embryonic stem cells as the treatment: a preliminary study

作　　者：瞿虎[1,2] 刘贵华[2] 袁浩锋 汪中扬[1] 杨轶[1] 张靖[2] 

机构地区：[1]中山大学附属第六医院泌尿外科,广东广州510655 [2]中山大学附属第六医院生殖中心,广东广州510655 [3]东莞市第三人民医院泌尿外科,广东东莞523326

出　　处：《广州医科大学学报》2015年第3期8-12,共5页Academic Journal of Guangzhou Medical University

摘　　要：目的:探讨大鼠隐睾模型的建立和胚胎干细胞睾丸内原位注射对热损伤所致的睾丸生精功能损害的疗效。方法:Wistar大鼠20只,取右睾丸手术建立隐睾模型,于7、10、14、21及28 d后行隐睾下降固定术,下降后选取不同的时间点行附睾穿刺液及睾丸病理检查;另Wistar大鼠20只,随机分为A组(n=10)和B组(n=10)。A组左睾丸为干细胞组(ES),右睾丸为自身空白对照(NPBS);B组左睾丸为组间对照组(JPBS)。各组在隐睾建模2周后行隐睾下降固定术并同时行睾丸内原位注射处理,2周后比较生精小管的直径(d)、精小管百分率(r)和生精小管的Johnsen评分(z)。结果:隐睾建模后10、14、21、30 d及隐睾复降后2月的附睾穿刺液中均未能发现精子,睾丸复降的中后期,部分生精小管的生精细胞层数仍然存在不同程度的恢复,含有生精细胞的d继续增加(P<0.05);干细胞注射后各组附睾穿刺液内均未见精子,ES组与NPBS组生精小管z差异有统计学意义(P<0.05)。结论:短期的睾丸热损伤是可逆的,超过10 d则严重影响到睾丸的生精功能,原位注射胚胎干细胞在本实验中尚不能证实对睾丸损伤有修复作用,但此研究方向仍具有极高的可行性。Objective： To investigate the effect of orthotopic injection of embryonic stem（ ES） cells as a treatment for impaired spermatogenesis of the testes caused by thermal damage in a rat model of cryptorchidism.Methods： Unilateral cryptorchidism in the right testis was surgically induced in 20 Wistar rats. Orchidopexy was performed on days 7,10,14,21 and 28. Epididymis puncture and testicular biopsy were done at different time points. Another 20 rats were randomly divided into groups A and B（ n = 10 each）. For group A,ES cells were injected in the left testis（ group ES） with the right testis as interclass control（ group NPBS）. For group B,the left testis in group B was intraclass control（ group JPBS）. Orchidopexy were performed at 2 weeks after surgical cryptorchidism in 3 groups,and meanwhile,orthotopic ES cell injection was given. Then epididymis puncture and testicular biopsy were done 2 weeks later. The diameter（ d）,percentage（ r） and Johnsen score（ z） of seminiferous tubules were compared in 3 groups.Results： No sperm was found in the epididymis puncture fluid on days 10,14,21 and 30 after surgical cryptorchidism and at 2 months after orchidopexy. However,certain recovery of seminiferous tubule structure remained at the mid-to-late time points after orchidopexy,along with increase in diameter of seminiferous tubules（ P〈0.05）. No sperm was found in the epididymis puncture fluid after ES cells injection. But the seminiferous Johnsen scores were significantly different between group ES and group NPBS（ P〈0.05）.Conclusion： While short term thermal damage of the testis is reversible,such damage lasting over 10 days could seriously damage spermatogenesis function. Although orthotopic injection of ES cells did not appear to help repair the testicle damage during our study period,the feasibility of our attempt remains promising.

关 键 词：睾丸 热损伤 胚胎干细胞 原位注射 

分 类 号：R697.22[医药卫生—泌尿科学]