己烯雌酚对小鼠睾丸引带中LGR8表达的影响  

Diethylstilbestrol affects LGR8 expression in mouse gubernaculum testis

作　　者：杨新彬[1,2,3] 蒋学武[1,2] 段守兴[1,2] 祁艳卫[1,2] 张镟[1,2] 李建宏[1,2] 

机构地区：[1]汕头大学医学院 [2]汕头大学医学院第二附属医院小儿外科,广东汕头515041 [3]广东省中医院普通外科,广东广州510120

出　　处：《中华男科学杂志》2012年第8期681-686,共6页National Journal of Andrology

基　　金：国家自然科学基金(81170555);广东省自然科学基金(9151009001000021)~~

摘　　要：目的:通过体内实验研究己烯雌酚(DES)对小鼠睾丸引带中胰岛素样因子3受体LGR8的影响,从而探讨外源性雌激素对小鼠睾丸下降的影响。方法:8～10周龄KM孕鼠随机分为正常组、空白对照组和实验组(0.1、1.0、10、100μg/kg DES 4个剂量组),共6组,每组20只。于孕9～17 d每天分别给予实验组妊娠小鼠不同剂量的DES(0.1、1、10、100μg/kg),空白对照组给予等体积DMSO+生理盐水,正常组不给药。应用免疫组化和RT-PCR分别检测胎鼠和幼鼠睾丸引带中LGR8蛋白和mRNA的表达。结果:HE染色可见胎鼠正常组、对照组睾丸引带发育良好,中间的间叶组织和外周的肌源细胞之间分界清楚;实验组可见睾丸引带发育不良,间叶组织和肌源细胞之间无明显分界,组织结构紊乱。幼鼠正常组和实验组睾丸引带发育未见明显不同。LGR8表达于睾丸引带肌源细胞和间叶组织细胞,以肌源细胞表达为主。实验组LGR8阳性表达较正常组弱,胎鼠各实验组和幼鼠DES 1、10、100μg组与同发育阶段的正常组相比均有统计学意义(P<0.05)。DES高剂量(10、100μg)组与同发育阶段的正常组相比LGR8 mRNA表达增加(P<0.05)。各实验组PCR产物测序均未见明显突变。结论:DES可影响小鼠睾丸引带LGR8 mRNA和蛋白的表达,DES可能通过影响INSL3-LGR8信号系统干扰睾丸引带的发育,进而影响睾丸正常下降。Objective： To investigate the impact of prenatal exposure to diethylstilbestrol （DES） on the specific receptor LGR8 of insulin-like factor 3 （ INSL3 ） in the mouse gubernaculum testis, and that of exoestrogens on deseensus testis in mice. Methods ： A total of 120 pregnant KM mice aged 8 to 10 weeks were assigned to a normal, a blank control and 4 DES groups of equal number, the blank controls injected subcutaneously with dimethyl sulfoxide plus normal saline, and the DES groups with DES at 0. 1, 1, 10 and 100 μg,/kg body weight, respectively, from embryonic day 9 （ED9） through ED17. Immunohistochemistry and RT-PCR were used to detect the expressions of LGR8 protein and mRNA in the gnbernacula testis of the ED18 fetuses and PND20 （postnatal day 20 ） off- spring of the mice. Results ： Histological analysis showed that the gubernacula testis of the ED18 fetuses were well developed in both the normal and control groups, with an inner mesenchymal core and muscular outer layer. In contrast, the gubernacula testis were poorly developed in the experimental groups, morphologically abnormal and without visible dividing line between the mesenchymal tissue and the muscular outer layer. No obvious differences were found in the gubemaculum testis development of the neonates between the normal and experimental groups. Positive immunostaining was seen in the mesenchymal core and muscular outer layer, but mainly in the latter. The expression of LGR8 was weaker in the experimental groups than in the normal group （P 〈 0.05 ）, but that of LGR8 mRNA was increased in the high-dose （10 and 100 μg/kg） DES groups （P 〈 0.05 ）. No obvious mutations were observed in the PCR products in any of the experimental groups. Conclusion： Prenatal exposure to diethylstilbestrol affected the expression of LGR8 mRNA in the mouse gubemaculum testis, which suggests that diethylstilbestrol may induce cryptorchidism by interfering with the INSL3-LGR8 signaling system and consequently the development of the gubemacu

关 键 词：己烯雌酚 睾丸引带 富含亮氨酸的G蛋白偶联受体 胰岛素样因子3 

分 类 号：R697.22[医药卫生—泌尿科学]