邻苯二甲酸二_论文.docxVIP

邻苯二甲酸二 作者：宋晓峰，魏光辉，邓永继，张德迎，陈旋，刘星 【关键词】 二甲酸二 【Abstract】 AIM: To explore the mRNA expression of INSL3 gene in Leydig cells of fetal and neonatal mice exposed to di(2ethylhexyl)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 betahydroxysteroid dehydrogenase (3βHSD). Reverse transcriptasepolymerase chain reaction (RTPCR) 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 RTPCR. 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 offsprings testis were lower than those of the control groups. CONCLUSION: DEHP can downregulate the expression level of INSL3 mRNA in Leydig cells in vitro and in vivo. It may be a possible mechanism that DEHP interferes with gubernaculum development and causes cryptorchidism. 【Keywords】 Di(2ethylhexyl), phthalate；fetus；Leydig cell；in situ hybridization ；RTPCR 【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：DEHP 50，100，200 mg/L分别作用于原代培养的小鼠胚胎Leydig细胞，应用RTPCR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500 mg/kg?d）灌胃作用于GD（Gestation day）12～PND（Postnatal day）3孕鼠，RTPCR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 【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 0引言 邻苯二甲酸二（2乙基）己酯（DEHP），是塑料工业中广泛使用的增塑剂，是邻苯二甲酸酯类化合物（PAE）中的一种，也是公认的环境内分泌干扰物，可影响机体内分泌