T Zhang1, C Zhao2, L Luo3, J Xiang4, J Cheng4, T Wang4, D Chen5. 1. Wuxi Maternity and Child Health Hospital Affiliated to Nanjing Medical University, Wuxi 214002, China. Electronic address: zhangting040715@163.com. 2. State Key Laboratory of Reproductive Medicine, Nanjing Maternity and Child Health Hospital Affiliated to Nanjing Medical University, Nanjing 210004, China. 3. Department of Respiratory Medicine, Jinling Hospital, Nanjing University Medical School, Nanjing 210093, China; Wuxi Second Hospital Affiliated to Nanjing Medical University, Wuxi 214002, China. 4. Wuxi Maternity and Child Health Hospital Affiliated to Nanjing Medical University, Wuxi 214002, China. 5. Wuxi Maternity and Child Health Hospital Affiliated to Nanjing Medical University, Wuxi 214002, China. Electronic address: chendaozhen@163.com.
Abstract
INTRODUCTION: Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy-specific disease associated with a significant risk of fetal complications. Our previous study using an iTRAQ-based proteomics approach showed that ERp29 was overexpressed in the placenta tissue of ICP patients, which was an apoptosis-related protein and has not been investigated in the pathogenesis of ICP. The aim of this study was to explore the role of ERp29 in the mechanism of apoptosis in the placenta of ICP. METHODS: HTR-8/SVneo cells were cultured and treated with different concentrations of taurocholic acid (TCA) (0, 10, 50 and 100 μM). The apoptotic index and cell cycle were detected by flow cytometry; furthermore, the expression levels of ERp29 and p-p38 were detected by western blot. The ERp29-siRNA was also used to confirm the role of ERp29 in TCA induced-apoptosis. RESULTS: ERp29 expression and the apoptotic index were significantly increased in HTR-8/SVneo cells exposed to 100 μM TCA; so were p-p38 and caspase-3 activity, compared with the 50 μM, 10 μM TCA groups and negative control group (P < 0.05, respectively). The induction of apoptosis by TCA and the expression of p-p38 were reduced in HTR-8/SVneo cells after treatment with ERp29-siRNA, compared with controls (P < 0.05, respectively). CONCLUSIONS: This study suggested that overexpression of ERp29 may play a key role in TCA-induced apoptosis in HTR-8/SVneo cells via activation of p38, which may participate in the pathogenesis of ICP and may represent a novel target for ICP treatment.
INTRODUCTION:Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy-specific disease associated with a significant risk of fetal complications. Our previous study using an iTRAQ-based proteomics approach showed that ERp29 was overexpressed in the placenta tissue of ICP patients, which was an apoptosis-related protein and has not been investigated in the pathogenesis of ICP. The aim of this study was to explore the role of ERp29 in the mechanism of apoptosis in the placenta of ICP. METHODS: HTR-8/SVneo cells were cultured and treated with different concentrations of taurocholic acid (TCA) (0, 10, 50 and 100 μM). The apoptotic index and cell cycle were detected by flow cytometry; furthermore, the expression levels of ERp29 and p-p38 were detected by western blot. The ERp29-siRNA was also used to confirm the role of ERp29 in TCA induced-apoptosis. RESULTS:ERp29 expression and the apoptotic index were significantly increased in HTR-8/SVneo cells exposed to 100 μM TCA; so were p-p38 and caspase-3 activity, compared with the 50 μM, 10 μM TCA groups and negative control group (P < 0.05, respectively). The induction of apoptosis by TCA and the expression of p-p38 were reduced in HTR-8/SVneo cells after treatment with ERp29-siRNA, compared with controls (P < 0.05, respectively). CONCLUSIONS: This study suggested that overexpression of ERp29 may play a key role in TCA-induced apoptosis in HTR-8/SVneo cells via activation of p38, which may participate in the pathogenesis of ICP and may represent a novel target for ICP treatment.
Authors: Yun-Hui Zhang; Visar Belegu; Yu Zou; Fang Wang; Bao-Jiang Qian; Ran Liu; Ping Dai; Wei Zhao; Fa-Bao Gao; Lei Wang; Li-Mei Cao; John W McDonald; Su Liu; Na Lin; Ting-Hua Wang Journal: Mol Neurobiol Date: 2014-09-10 Impact factor: 5.590