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O1012 AQP3-null mice – Oligohydramnios and FGR model
Free communication (oral) presentations / International Journal of Gynecology & Obstetrics 107S2 (2009) S93–S396
O1010 Bile acid transporters expression profile in human placenta with intrahepatic cholestasis of pregnancy A. Xing1 , L. Wu1 , X. Yang. 1 Department of OB/GYN, Second Hospital of Sichuan University Objectives: Intrahepatic cholestasis of pregnancy (ICP) is a condition with increased fetal morbidity and mortality. There is evidence suggested that the poor fetal outcome is relating to fetal cholestasis. We supposed disturbed bile acid transport across the placenta is a potential reason. This study was designed to investigate the bile acid transporter expression profile in human placenta, and preliminarily evaluate the hepatobiliary-like excretory function of placenta in the pathology of fetal cholestasis with ICP. Materials and Methods: Placentas were collected from 20 normal pregnancy (control group) and 20 ICP (ICP group). Real time RTPCR was applied to detect the mRNA of bile acid transporters: OATP-A, OATP-C, MRP1, MRP2, FIC1, MDR3, BSEP and AE2. The basal membrane (BM) and microvillous membrane (MVM) of trophoblast were prepared with Illsley’s method. Protein expression of each transporter on membranes was measured by Western Blots. Comparisons between two groups were made by the independent samples t-test or two-independent samples test. A value of P < 0.05 was considered significant. Results: (1) All the transcripts except OATP-C and BSEP were detected in human placenta. OATP-A and MRP1 were located on the BM, while MRP2, FIC1, MDR3 and AE2 on the MVM; (2) OATP-A, AE2 gene expression were higher while FIC1 mRNA expression was lower in ICP than those in control group (P < 0.05). There were no significant differences among other genes between two groups. Conclusions: The expression pattern of bile acid transporters in placenta is different from that in liver, and its diversification appears to suggest that disturbed bile acid transporter system in placenta is involved in the pathological mechanism of fetal cholestasis in ICP. O1011 AQPs expression and regulation at human placenta and fetal membranes Z. Xiong, H. Liu, X. Song, R. Hao. Dept of Obstetrics, Third Affiliated Hospital of Guangzhou Medical College Objectives: To detect AQP1, 3, 8, 9 expression and regulation at human placenta and fetal membranes. Method: Placenta and fetal membranes were collected from women who presented with oligohydramnios (5 cases), polyhydramnios (5 cases), normal amniotic fluid volume (5 cases) underwent elective cesarean sections at term. Expression and localization of AQPs (AQP1, 3, 8, 9) was detected by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot analysis and immunohistochemistry. Results: AQP1, 3, 8, 9 were found their location at human placenta and fetal membranes, respectively. Furthermore, RT-PCR showed that the expression of AQP1 mRNA were significantly lower in oligohydramnios placenta and fetal membranes than normal pregnancy placenta and fetal membranes at term (P = 0.049, P = 0.022); the expression of AQP8 mRNA were significantly lower in oligohydramnios groups than that of normal pregnancy placenta at term (P = 0.037). The expression of AQP9 mRNA were significantly higher in polyhydramnios groups than that of normal pregnancy fetal membranes at term (P = 0.002). Conclusion: These findings suggested that AQPs expression and regulation at human placenta and fetal membranes may play important roles in maternal-fetal homeostasis and amniotic fluid balance.
S381
O1012 AQP3-null mice – Oligohydramnios and FGR model Z. Xiong, H. Liu, Z. Zheng, T. Ma. Dept of Obstetrics, Third Affiliated Hospital of Guangzhou Medical College, Guangzhou, China Membrane Channel research laboratory, Northeast Normal University, Changchun, China Objective: The cell membrane water channel protein aquaporin 3 (AQP3) may play an important role in the homeostasis of maternalfetal fluid, and also in pregnancy. But there is no direct evidence for that. AQP3 knockout mice are very useful animal model for research AQP3 function in vivo. The objective of the present study was to investigate the role of AQP3 in the homeostasis of maternal-fetal fluid during pregnancy by AQP3 null mice. Method: Homozygous AQP3 knockout mice were mated. The pregnancy AQP3 knockout mice were divided into 5 subgroups (e7, e13, e16, e18 and full term) depended on gestational age, the wild-type (KM mice) pregnancy mice as the control group. Total embryo and atrophies embryo numbers were recorded in each subgroup. Fetus weight, placenta weight, placenta area, placenta water content and the amniotic fluid were measured at e13, e16, e18, and amniotic fluid at 16GD was collected to detection the composition and osmolality of amniotic fluid. A one-way analysis of variance on ranks and chi-square test were used for statistics. Results: Compared to their wild-type mice, the number of AQP3-KO pregnant mice embryo was much lesser than that in KM pregnant mice (P = 0.001) at the early trimester of pregnancy (e7), and significant decrease was found in AQP3 null pregnant mice with the development of gestational age. The fetal mice weight of AQP3 null pregnancy mice was significant lighter than that of the wild type (KM mice) at e16 and e18. That is to say, fetal growth restriction (FGR) accurred in AQP3 null mice. The placenta area of AQP3-KO mice is significant bigger than their wild type during different gestational ages. The placenta weight of AQP3-KO is significant heaver than their wild type at e13 and e16, however there was no difference between the two groups at e18. The placenta water content of AQP3-KO is more than that of KM placenta at e13. Amniotic fluid volume of AQP3-KO mice is significant decrease compare to their wild type at e18, through there was no difference between the two groups at e13 and e16. The AQP3 null mice had a higher Urea, Creat, Ka, Na, Cl than their wide-type mice, and also have a higher osmolality of amniotic fluid. Conclusions: This study provides the evidences that AQP3 may play an important role in homeostasis of maternal-fetal fluid and also in pregnancy. Acknowledgement: Thanks to Dr Verkman for providing AQP3 knockout mice and his suggestions at our research. O1013 Expression of NKG2D and its ligands in pelvic endometriosis H. Xu, Q.D. Lin, Z.P. Cheng, Q.L. Ma, X.P. Wang Background: To investigate the expression of NKG2D, an importmant activating receptor, on NK cells in peripheral blood and peritoneal fluid with or without endometriosis and the expression of its ligands, MICA/MICB and ULBP-1, 2, 3, 4, on eutopic and ectopic endometrial cells with pelvic endometriosis and eutopic endometrial cells without endometriosis. Methods: NKG2D on NK cells in peripheral blood and peritoneal fluid of 20 patients with pelvic endometriosis and 13 women without endometriosis were examined with flow cytometry. The expression of the ligands of NKG2D of ectopic endometrial cells (n = 27), eutopic endometrial cells with pelvic endometriosis (n = 14) and eutopic endometrial cells without endometriosis (n = 15) was determined with real time PCR and Western Blotting simultaneously. All subjects were accepted operation in Renji Hospital and Yang Pu Central Hospital between February 2006 and October 2006.
O1010 Bile acid transporters expression profile in human placenta with intrahepatic cholestasis of pregnancy A. Xing1 , L. Wu1 , X. Yang. 1 Department of OB/GYN, Second Hospital of Sichuan University Objectives: Intrahepatic cholestasis of pregnancy (ICP) is a condition with increased fetal morbidity and mortality. There is evidence suggested that the poor fetal outcome is relating to fetal cholestasis. We supposed disturbed bile acid transport across the placenta is a potential reason. This study was designed to investigate the bile acid transporter expression profile in human placenta, and preliminarily evaluate the hepatobiliary-like excretory function of placenta in the pathology of fetal cholestasis with ICP. Materials and Methods: Placentas were collected from 20 normal pregnancy (control group) and 20 ICP (ICP group). Real time RTPCR was applied to detect the mRNA of bile acid transporters: OATP-A, OATP-C, MRP1, MRP2, FIC1, MDR3, BSEP and AE2. The basal membrane (BM) and microvillous membrane (MVM) of trophoblast were prepared with Illsley’s method. Protein expression of each transporter on membranes was measured by Western Blots. Comparisons between two groups were made by the independent samples t-test or two-independent samples test. A value of P < 0.05 was considered significant. Results: (1) All the transcripts except OATP-C and BSEP were detected in human placenta. OATP-A and MRP1 were located on the BM, while MRP2, FIC1, MDR3 and AE2 on the MVM; (2) OATP-A, AE2 gene expression were higher while FIC1 mRNA expression was lower in ICP than those in control group (P < 0.05). There were no significant differences among other genes between two groups. Conclusions: The expression pattern of bile acid transporters in placenta is different from that in liver, and its diversification appears to suggest that disturbed bile acid transporter system in placenta is involved in the pathological mechanism of fetal cholestasis in ICP. O1011 AQPs expression and regulation at human placenta and fetal membranes Z. Xiong, H. Liu, X. Song, R. Hao. Dept of Obstetrics, Third Affiliated Hospital of Guangzhou Medical College Objectives: To detect AQP1, 3, 8, 9 expression and regulation at human placenta and fetal membranes. Method: Placenta and fetal membranes were collected from women who presented with oligohydramnios (5 cases), polyhydramnios (5 cases), normal amniotic fluid volume (5 cases) underwent elective cesarean sections at term. Expression and localization of AQPs (AQP1, 3, 8, 9) was detected by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot analysis and immunohistochemistry. Results: AQP1, 3, 8, 9 were found their location at human placenta and fetal membranes, respectively. Furthermore, RT-PCR showed that the expression of AQP1 mRNA were significantly lower in oligohydramnios placenta and fetal membranes than normal pregnancy placenta and fetal membranes at term (P = 0.049, P = 0.022); the expression of AQP8 mRNA were significantly lower in oligohydramnios groups than that of normal pregnancy placenta at term (P = 0.037). The expression of AQP9 mRNA were significantly higher in polyhydramnios groups than that of normal pregnancy fetal membranes at term (P = 0.002). Conclusion: These findings suggested that AQPs expression and regulation at human placenta and fetal membranes may play important roles in maternal-fetal homeostasis and amniotic fluid balance.
S381
O1012 AQP3-null mice – Oligohydramnios and FGR model Z. Xiong, H. Liu, Z. Zheng, T. Ma. Dept of Obstetrics, Third Affiliated Hospital of Guangzhou Medical College, Guangzhou, China Membrane Channel research laboratory, Northeast Normal University, Changchun, China Objective: The cell membrane water channel protein aquaporin 3 (AQP3) may play an important role in the homeostasis of maternalfetal fluid, and also in pregnancy. But there is no direct evidence for that. AQP3 knockout mice are very useful animal model for research AQP3 function in vivo. The objective of the present study was to investigate the role of AQP3 in the homeostasis of maternal-fetal fluid during pregnancy by AQP3 null mice. Method: Homozygous AQP3 knockout mice were mated. The pregnancy AQP3 knockout mice were divided into 5 subgroups (e7, e13, e16, e18 and full term) depended on gestational age, the wild-type (KM mice) pregnancy mice as the control group. Total embryo and atrophies embryo numbers were recorded in each subgroup. Fetus weight, placenta weight, placenta area, placenta water content and the amniotic fluid were measured at e13, e16, e18, and amniotic fluid at 16GD was collected to detection the composition and osmolality of amniotic fluid. A one-way analysis of variance on ranks and chi-square test were used for statistics. Results: Compared to their wild-type mice, the number of AQP3-KO pregnant mice embryo was much lesser than that in KM pregnant mice (P = 0.001) at the early trimester of pregnancy (e7), and significant decrease was found in AQP3 null pregnant mice with the development of gestational age. The fetal mice weight of AQP3 null pregnancy mice was significant lighter than that of the wild type (KM mice) at e16 and e18. That is to say, fetal growth restriction (FGR) accurred in AQP3 null mice. The placenta area of AQP3-KO mice is significant bigger than their wild type during different gestational ages. The placenta weight of AQP3-KO is significant heaver than their wild type at e13 and e16, however there was no difference between the two groups at e18. The placenta water content of AQP3-KO is more than that of KM placenta at e13. Amniotic fluid volume of AQP3-KO mice is significant decrease compare to their wild type at e18, through there was no difference between the two groups at e13 and e16. The AQP3 null mice had a higher Urea, Creat, Ka, Na, Cl than their wide-type mice, and also have a higher osmolality of amniotic fluid. Conclusions: This study provides the evidences that AQP3 may play an important role in homeostasis of maternal-fetal fluid and also in pregnancy. Acknowledgement: Thanks to Dr Verkman for providing AQP3 knockout mice and his suggestions at our research. O1013 Expression of NKG2D and its ligands in pelvic endometriosis H. Xu, Q.D. Lin, Z.P. Cheng, Q.L. Ma, X.P. Wang Background: To investigate the expression of NKG2D, an importmant activating receptor, on NK cells in peripheral blood and peritoneal fluid with or without endometriosis and the expression of its ligands, MICA/MICB and ULBP-1, 2, 3, 4, on eutopic and ectopic endometrial cells with pelvic endometriosis and eutopic endometrial cells without endometriosis. Methods: NKG2D on NK cells in peripheral blood and peritoneal fluid of 20 patients with pelvic endometriosis and 13 women without endometriosis were examined with flow cytometry. The expression of the ligands of NKG2D of ectopic endometrial cells (n = 27), eutopic endometrial cells with pelvic endometriosis (n = 14) and eutopic endometrial cells without endometriosis (n = 15) was determined with real time PCR and Western Blotting simultaneously. All subjects were accepted operation in Renji Hospital and Yang Pu Central Hospital between February 2006 and October 2006.