邻苯二甲酸二(2-乙基)己酯对胚胎及新生小鼠睾丸Leydig细胞INSL3 mRNA表达的影响  

Effects of di(2-ethylhexyl)phthalate on mRNA expression of INSL3 gene in leydig cells in fetal and neonatal mice

作　　者：宋晓峰[1] 魏光辉[1] 邓永继[1] 张德迎[1] 陈旋[1] 刘星[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：《第四军医大学学报》2006年第21期1973-1976,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30371475)

摘　　要：目的：通过体内体外实验探讨邻苯二甲酸二（2-乙基）己酯（DEHP）对胚胎及新生小鼠睾丸Leydig细胞INSL3 mRNA表达的影响．方法：DEHP50，100，200mg／L分别作用于原代培养的小鼠胚胎Leydig细胞，应用RT—PCR和原位分子杂交技术检测DEHP对Leydig细胞INSL3 mRNA表达的影响；DEHP分别以低、中、高三组剂量（100，200和500mg／kg·d）灌胃作用于GD（Gestationday）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．结果：DEHP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；各实验组包括原代Leydig细胞及不同时间点的新生鼠睾丸INSL3 mRNA表达水平、表达相对强度均明显低于对照组（P〈0．05，或P〈0．01）．结论：DEHP下调小鼠睾丸Leydig细胞INSL3 mRNA的表达，可能是影响引带发育导致隐睾的机制之一．AIM： To explore the mRNA expression of INSL3 gene in Leydig cells of fetal and neonatal mice exposed to di （2- ethylhexyl） phthalate （DEHP） 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 （ 3 β-HSD ）. Reverse transcriptase-polymerase chain reaction （RT-PCR） and in situ hybridization were used to examine the expression levels of INSL3 mRNA in primarily cultured Leydig cells incubated with DEHP （at the dose of 50, 100,200 mg/L, respectively）. Pregnant mice were exposed to DEHP at the dose of 100, 200 or 500 mg/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 cells and neonatal mouse testis exposed to DEHP were observed. The expression level of INSL3 mRNA in DEHP treated groups including the primarily cultured Leydig cells and male mouse offspring＇s testis were lower than those of the control groups. CONCLUSION： DEHP can downregnlate the expression level of INSL3 mRNA in Leydig cells in vitro and in vivo. It may be a possible mechanism that DEHP interferes with gnbernaculum development and causes cryptorchidism.

关 键 词：2-乙基己基 邻苯二甲酸 胚胎 LEYDIG细胞 原位分子杂交 逆转录聚合酶链反应 

分 类 号：TQ414.1[化学工程]