Yue Zhou1,2, Fangyuan Huang1, Yang Liu1, Dian Li1, Yu Zhou1, Lianju Shen1, Chunlan Long1, Xing Liu3, Guanghui Wei1. 1. Department of Pediatric Urology Surgery, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, 400014, Chongqing, China. 2. Chengdu Women's and Children's Central Hospital, School of Medicine, University of Electronic Science and Technology of China, 611731, Chengdu, China. 3. Department of Pediatric Urology Surgery, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, 400014, Chongqing, China. liux_0217@163.com.
Abstract
BACKGROUNDS: To investigate the potential mechanism of hypospadias induced by DEHP in rats to reveal the preventative effect of TGF-β1 in hypospadias induced by DEHP via the reduction of EMT. METHODS: Time-mated Sprague-Dawley rats underwent cesarean section, and the penises of male pups were collected after exposure to corn oil or DEHP to establish a rat model of hypospadias and to further study the molecular mechanisms of hypospadias in vivo. In addition, the penises were cultured and treated with MEHP or MEHP+TGF-β1 in vitro. Subsequently, histomorphology and elements in TGF-β/Smad signaling pathway changes were evaluated using scanning electron microscopy, immunofluorescence, polymerase chain reaction, and western blot. RESULTS: The development of rat penis and urethral seam fusion were delayed after the treatment with DEHP in vivo or MEHP in vitro compared with the Control group. Moreover, TGF-β1, Smad2/Smad3, and the mesenchymal biomarkers, including α-SMA, N-cadherin, and Vimentin, were decreased. However, the epithelial biomarkers, including E-cadherin, ZO-1, β-catenin, and occludin, were increased. In addition, TGF-β1 could relieve all of the above changes. CONCLUSION: Gestational DEHP exposure could lead to hypospadias by reducing urethral EMT. Moreover, TGF-β1 could prevent it by regenerating EMT through activating the TGF-β/Smad signal pathway.
BACKGROUNDS: To investigate the potential mechanism of hypospadias induced by DEHP in rats to reveal the preventative effect of TGF-β1 in hypospadias induced by DEHP via the reduction of EMT. METHODS: Time-mated Sprague-Dawley rats underwent cesarean section, and the penises of male pups were collected after exposure to corn oil or DEHP to establish a rat model of hypospadias and to further study the molecular mechanisms of hypospadias in vivo. In addition, the penises were cultured and treated with MEHP or MEHP+TGF-β1 in vitro. Subsequently, histomorphology and elements in TGF-β/Smad signaling pathway changes were evaluated using scanning electron microscopy, immunofluorescence, polymerase chain reaction, and western blot. RESULTS: The development of rat penis and urethral seam fusion were delayed after the treatment with DEHP in vivo or MEHP in vitro compared with the Control group. Moreover, TGF-β1, Smad2/Smad3, and the mesenchymal biomarkers, including α-SMA, N-cadherin, and Vimentin, were decreased. However, the epithelial biomarkers, including E-cadherin, ZO-1, β-catenin, and occludin, were increased. In addition, TGF-β1 could relieve all of the above changes. CONCLUSION: Gestational DEHP exposure could lead to hypospadias by reducing urethral EMT. Moreover, TGF-β1 could prevent it by regenerating EMT through activating the TGF-β/Smad signal pathway.