Phthalate.doc

 PAGE \* MERGEFORMAT 2 Phthalate Authors: Xiao-Feng Song, Guang-Hui Wei, DENG Yong-Ji, Zhang Ying, Chen Xuan, Liu Xing [Keywords:] dimethyl [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 (3β HSD). 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 [Abstract] Objective: To explore the in vivo experiments in vitro phthalate Bis (2-ethyl) hexyl (DEHP) on the embryonic and neonatal mouse testis INSL3 mRNA expression in Leydig cells. Methods: DEHP 50,100,200 mg / L respectively, acting on the primary culture of mouse embryonic Leydig cells, application of RTPCR and in situ hybridization detection of DEHP on Leydig cells, INSL3 mRNA expression; DEHP, respectively