己烯雌酚干预下围产期小鼠睾丸Leydig细胞INSL3 mRNA的表达变化  

The change of the mRNA expression of INSL3 gene in perinatal mice leydig cells exposed to diethylstilbestrol in vivo and in vitro

作　　者：宋晓峰[1] 张德迎[1] 刘星[1] 吴盛德[1] 邓永继[1] 魏光辉[1] 

机构地区：[1]重庆医科大学附属儿童医院泌尿外科,重庆400014

出　　处：《重庆医科大学学报》2009年第5期513-516,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金委资助项目(30371475)。

摘　　要：目的:通过体内、体外实验研究己烯雌酚(Diethylstilbestrol,DES)对KM小鼠围产期小鼠睾丸Leydig细胞胰岛素样因子3(Insuliu-like hormone3,INSL3)mRNA表达的影响,探讨隐睾发生的可能机制。方法:DES分别以低、中、高3组剂量(1、5、25μg/L)分别作用于原代培养的胎龄16d小鼠胚胎Leydig细胞,应用RT-PCR和原位分子杂交技术检测DES对睾丸Leydig细胞INSL3mRNA的表达水平;DES分别以低、中、高3组剂量[1μg(/kg·d)、10μg(/kg·d)、100μg(/kg·d)]灌胃作用于GD(Gestation day)12～PND(Postnatal day)3孕鼠,RT-PCR检测新生小鼠睾丸INSL3 mRNA表达的相对强度,并比较其差异性。结果:DES改变原代培养的小鼠胚胎Leydig细胞和新生小鼠睾丸的形态结构,以中、高剂量组改变尤为明显;体外实验中,DES中、高剂量组原代Leydig细胞INSL3 mRNA表达水平均明显低于对照组(P<0.01)和DES低剂量组(P<0.01);DES试验组Leydig细胞INSL3 mRNA表达相对强度低于对照组(P<0.01)。体内实验中,DES试验组不同时间点的新生鼠睾丸INSL3 mRNA表达相对强度均明显低于对照组(P<0.05,或P<0.01),实验组组间两两比较也有差异(P<0.05,或P<0.01)。结论:DES可下调睾丸Leydig细胞INSL3 mRNA的表达水平,其可能是影响小鼠睾丸引带发育导致隐睾的机制之一。Objective： To explore the mRNA expression of INSL3 gene in perinatal mice Leydig cells exposed to diethylstilbestrol （ DES ） in vivo and in vitro. Methods： The Leydig cells were isolated from fetal mouse testis and were cultured in DMEM/F12 medium and identified by 3 beta-hydroxysteroid dehydrogenase（ 3β-HSD ）. Reverse transcriptase-polymerase chain reaction（ RT-PCR ）and in situ hybridization were used for examination of the expression levels of INSL3 mRNA in primarily cultured Leydig cells incubated with DES（at the dose of 1 μg/L,5 μg/L and 25 μ g/L,respectively）. Pregnant mice were exposed to DES at the dose of 1,10 or 100 μg/kg body weight per day by gavage from gestation day （GD）12 to postnatal day （PND）3.The expression of INSL3 gene in neonatal mouse testis was investigated by RT-PCR. Results： Histological alterations of primarily cultured Leydig ceils and neonatal mouse testis exposed to DES were observed. The expression level of INSL3 mRNA in DES treated groups including the primarily cultured Leydig cells and male mouse offspring＇s testis were lower than those of the control groups. Conclusion： DES can downregulate the expression level of INSL3 mRNA in Leydig ceils in vitro and in vivo. It may be a possible mechanism that DES interferes with gubernaculum development and cause cryptorchidism.

关 键 词：己烯雌酚 胚胎 LEYDIG细胞 原位分子杂交 逆转录聚合酶链反应 胰岛素样因子3 

分 类 号：R994.6[医药卫生—毒理学] Q492.4[医药卫生—药学]