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目的探讨新西兰大白兔睾丸皮下埋藏导致精子发生障碍后,将埋藏之睾丸放回正常阴囊内其精子发生障碍是否可逆。方法育龄期雄性新西兰大白兔42只,曾有过生育史的雌性12只,将雄兔应用随机数字表法完全随机分成3组:空白组(12只);双埋组(双侧睾丸埋藏组12只);单埋组(单侧睾丸埋藏组18只)。双埋组将睾丸埋藏于两侧腹股沟区皮下;单埋组将一侧睾丸置于腹股沟区皮下,另一侧睾丸保留在阴囊内;空白组不作任何处理。模型成功后第9周空白组、双埋组、单埋组分别随机取6只进行睾丸活检,均行HE染色及采用TUNEL法检测生精细胞凋亡情况,同时对空白组及双埋组的剩余6只与分别与雌兔一一配对喂养观察生育情况。同期将单埋组未取活检的12只动物的未处理侧睾丸从正常阴囊内移出埋藏至同侧腹股沟区皮下,而将前期埋藏之睾丸放回刚移走睾丸的对侧正常阴囊内。单埋组睾丸放回阴囊后第4周及第9周分别对放回阴囊之睾丸取6只取活检,HE染色及TUNEL法检测生精细胞凋亡情况。结果与空白组相比双埋组模型建立后第9周的HE染色见曲细精管内生精细胞明显减少,排列紊乱,管腔内未见精子;TUNEL法检测结果显示精原细胞也有明显凋亡征象,生精细胞凋亡指数(apoptotic index,AI)明显高于空白组(P<0.05);配对喂养雌兔均无生育,生育率明显低于空白组(P<0.05)。单埋组模型建立后第9周埋藏之睾丸活检HE染色结果与双埋组相似,TUNEL法检测结果生精细胞的凋亡指数与双埋组相比较差异无统计学意义。埋藏睾丸放回阴囊术后第9周的HE染色示曲细精管内生精细胞层数增多,可见成熟精子,凋亡指数与双埋组比较明显降低。结论睾丸皮下埋藏与皮瓣重建阴囊一样会导致睾丸精子发生障碍,但这种损害是可逆的。
Objective To investigate whether the spermatogenesis disorder of New Zealand white rabbit testis buried in the testis can lead to spermatogenesis disorder and put it back in the normal scrotum. METHODS: Forty-two male New Zealand white rabbits of childbearing age were randomly divided into three groups: blank group (n = 12), double-burial group Only); single buried group (unilateral testicular buried group of 18). In the double-burial group, the testes were buried under the inguinal region on both sides. In the single-burial group, one side of the testes was placed in the groin area and the other side of the testes in the scrotum. The blank group was not treated. At 9 weeks after the model was established, the blank group, the double-buried group and the single-buried group were randomly selected to perform the testicular biopsy. HE staining and TUNEL method were used to detect the apoptosis of spermatogenic cells. At the same time, The remaining 6 and females were paired with one by one feeding observed fertility. In the same period, untreated testes from 12 animals without biopsy were removed from the normal scrotum to the ipsilateral groin area, and the pre-buried testicles were placed back into the normal scrotum just opposite to the testes. Single testis into the scrotum after 4 weeks and 9 weeks, respectively, returned to the scrotum of the testis to take six biopsies, HE staining and TUNEL assay apoptosis of spermatogenic cells. Results Compared with the blank group, the HE staining at the 9th week after establishment of the double-buried group showed that the number of spermatogenic cells in the seminiferous tubules was significantly reduced, disordered and no spermatozoa were found in the lumen. TUNEL assay showed that the spermatogonia also showed obvious apoptosis The apoptotic index (AI) was significantly higher than that of the blank group (P <0.05). The female rabbits fed without mating had no fertility and the fertility rate was significantly lower than that of the blank group (P <0.05). The result of TUNEL showed that the apoptosis index of spermatogenic cells was not significantly different from that of double-burial group at the 9th week after establishment of single-burial model. Hematoxylin-eosin staining showed that the number of endogenous spermatocytes in the seminiferous tubules increased at the 9th week after the testis was put back into the scrotum. The apoptotic index of the mature spermatozoa was obviously decreased compared with that of the double burial group. Conclusion Subcutaneous testicular skin flap reconstruction and scrotum can lead to testicular spermatogenesis disorder, but this damage is reversible.