我们利用RNA体内干扰技术降低正常小鼠睾丸组织中冷诱导RNA结合蛋白(CIRP)蛋白的表达,观察RNA干扰后小鼠睾丸重量、生精小管形态学变化及生精细胞凋亡的改变,探讨CIRP与生精功能的关系. 一、材料与方法 1.实验动物:30只BALB/c雄性,鼠龄8周,体质量18 ～20 g. 2.动物模型的建立:(1)小干扰RNA(siRNA)干扰组:在体视显微镜下于睾丸网内注入30μl的siRNA序列和转染试剂混合物.(2)siRNA阴性对照组即将siRNA序列换作无意义的RNA序列,其他同上.(3)正常对照组:未作任何处理.
Objective To study the relationship between the expression of endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) with the damage of contralateral testis of unilateral cryptorchidism in the experimental rats. Methods Forty-five immature male Sprague-Dawley rats (aged 22 days) were randomly divided into the group A (unilateral cryptorchidism and the ipsilateral genitofemoral nerve division), group B (unilateral cryptorchidism), group C (sham operation), n = 15 in each group. When the rats were aged 65 days, all the rats were sacrificed and the testes were obtained. The morphological changes of the spermatogenic epithelium in the testes were observed, and the germ cell apoptosis was detected by the TdT-mediated dUTP-biotin nick end labeling (TUNEL) technique. The expression of eNOS and VEGF was detected by using Western blotting and immunohistochemistry in the testicular tissues.Results The germ cell apoptosis was increased significantly ( t1 = 3. 04, t2 = 3. 94, t1 ,t2 ＞ t28 , P ＜0. 01) , and the levels of eNOS and VEGF in the contralateral testes were also increased obviously in group B as compared with groups A and C ( P ＜ 0. 01) , but the entire indexes in groups A and C had no significant difference. Conclusion eNOS, VEGF and genitofemoral nerve play a important role in the damage of the contralateral testes of unilateral cryptorchidism, and the damage can be prevented by genitofemoral nerve division.
目的 研究大鼠隐睾睾丸组织生精功能,了解睾丸生殖细胞凋亡.方法 建立隐睾大鼠动物模型,获得单侧隐睾大鼠(A组)12只,双侧隐睾大鼠(B组)10只,空白对照正常大鼠(C组)10只.当日龄为60 d时,切取大鼠双侧睾丸组织,行HE染色、Johnsen评分;采用原位缺口末端标记法检测生殖细胞凋亡.结果 A、B组隐睾睾丸组织Johnsen评分较对照组显著降低(P<0.05),生精功能明显破坏,生殖细胞凋亡指数显著增加(P<0.05).结论大鼠隐睾睾丸组织生精功能受到损害,生殖细胞凋亡增加.
