GABA receptor A α3 subunit can be involved in the pathophysiology of preeclampsia

s / Placenta 35 (2014) A1eA23 A15 Methods: We performed ZnCP-Iimmunostaining in the 28 fetal membranes of term spontaneous vaginal delivery and 32 term elective cesarean section before initiation of labor between 2011 and 2012. Detection of meconium components was compared. Results: Heterogeneous patterns of positive immunostaining of ZnCPIwere observed in the chorionic and/or decidual tissues. Positive immunostaining of ZnCP-I was observed in all 28 fetal membranes of spontaneous vaginal deliveries (100%). By contrast, both positive (18 cases; 56%) and negative immunostaining of ZnCP-I (14 cases; 44%) were observed among 32 elective cesarean sections. Conclusion: It was suggested that the meconium may contribute to the imitation of labor as a signal of the fetal maturation. O-055. GABA RECEPTOR A a3 SUBUNIT CAN BE INVOLVED IN THE PATHOPHYSIOLOGY OF PREECLAMPSIA. Kaoru Kawasaki , Eiji Kondoh , Yoshitsugu Chigusa , Ryusuke Murakami , Hiroshi Takai , Mari Ujita , Hikaru Kiyokawa , Fumitomo Nishimura , Haruta Mogami , Satoru Takeda , Ikuo Konishi . Department of Obstetrics and Gynecology, Kyoto University Graduate School of Medicine, Japan; Department of Obstetrics and Gynecology, Jyuntendo University, Japan Objective: The receptor of Gamma-aminobutyric acid (GABA), an inhibitory neuro-transmitter, is expressed in placenta. However, the role of GABA receptor signalling in placenta has yet been fully clarified. The aim of the present study was to investigate the function of GARA receptor A a3 subunit (GABRA3) in placenta and its involvement in the pathophysiology of preeclampsia (PE). Methods: The expression of GABRA3 was examined in control and preeclamptic human placentas by RT-PCR and western blotting. The influence of oxidative stress, inflammation and hypoxia on GABRA3 was examined using immortalized human trophoblast cells (TCL1) treated with or without H2O2 (100~400microM), TNF-alpha (10ng/ml) and hypoxia (O2: 1%) for 24 hours. The effect of GABA A receptor agonist (Muscimol) on oxidative stress was evaluated by flow cytometric determination of reactive oxygen species (ROS) in TCL1 treated with or without H2O2 (100microM) for 1 hours. The influence of Muscimol on inflammation was investigated by western blotting of phospho-PKC and Nuclear factorkappaB (NF-kappaB) using TCL1 treated with or without TNF-alpha (10ng/ ml) for 30 minutes. Statistical comparisons were performed with unpaired Student t-test and one-way ANOVA followed by turkey’s multiple comparisons test. Results: The expression of GABRA3 was significantly decreased in the preeclamptic placenta. GABRA3 expressions in TCL1 were augmented by H2O2 and TNF-alpha (p<0.05, p1⁄40.0016, respectively). On the other hand, hypoxia decreased GABRA3 (p1⁄40.0048). Flow cytometry demonstrated that H2O2-induced ROS was increased by Muscimol (p<0.05). Moreover, the activation of phospho-PKC and nuclear translocation of NF-kappaB stimulated by TNF-alpha; were significantly increased by Muscimol, Conclusions: The activation of GABA receptor signaling was shown to increase the generation of ROS and inflammatory activity in trophoblast cells. These results suggested that decrease of GABRA3 in preeclamptic placenta may contribute to the protection of placenta. O-056. EXPRESSION OF EPITHELIAL-MESENCHYMAL TRANSITION-RELATED FACTORS IN PLACENTA ACCRETA Tokuro Shirakawa, Yoshiya Miyahara, Yasuhiko Ebina, Hideto Yamada. Department of Obstetrics and Gynecology, Kobe University Graduate School of Medicine, Japan Epithelial-mesenchymal transition (EMT) is a key process influencing cancer progression and metastasis. The purpose of this study was to investigate the expression of EMT-related factors in chorionic villi and decidual cells in placenta accreta. The current study included 19 patients diagnosed with placenta accreta after hysterectomy. The expression of E-cadherin, vimentin, Snail, and TGF-beta in placental tissues was analyzed by immunohistochemical staining. Immunostaining intensity was semi quantitatively evaluated using the HSCORE algorithm. In the chorionic villi of invasive placenta accreta tissues, Ecadherin expression was significantly lower than that in noninvasive tissues. In the decidual cells of the invasive placenta accreta tissues, expression of TGF-beta and Snail significantly increased. These results suggest that EMT may contribute to excessive trophoblast invasion into the myometrium in placenta accreta. O-057. A CASE OF CHORIOANGIOMA COMPLICATED WITH FETAL HEART FAILURE Miho Matsushima, Kei Tanaka, Yukiko Matsuzawa, Tomoko Izawa, Keiji Sakai, Mitsutoshi Iwashita. Department of Obstetrics and Gynecology,Kyorin University School of Medicine, Japan Chorioangiomas are recognized with the highest frequency in benign tumor of placenta, and a large size of chorioangioma is associated with several fetal complications such as fetal anemia, polyhydroamnios, cardiomegaly, hydrops fetalis, and fetal growth restriction. We report a case of giant chorioangioma complicated with hydrops fetalis followed by fetal cardiomegaly. A 38-year-old woman, gravida 1, was refered to our Department at 27 weeks gestational age (GA) because of a pregnancy of blood type incompatibility. At this time, estimated fetal body weights was 1,037g (-0.3SD) with no abnormal findings. At the 28 weeks GA, anti-Dimmunoglobulin was given her because of negative indirect Coombs test. At the 30 weeks GA, ultrasonography showed fetal cardiomegaly, cardiothoracic area ratio (CTAR) 42%. Though we suspected that caused by congenital cardiac anomaly and blood type incompatible pregnancy, we could not identify the cause. At the 31weeks GA, the fetus showed hydrops fetalis with skin edema, pericardial effusion, CTAR 44%, and elevation of inferior vena cava preload index. Color Doppler ultrasonography showed a hypervascular tumor (81 X 95cm) in placenta. Cesarean section was performed due to diagnosis with fetal heart failure from giant chorioangioma at 32 weeks GA. The newbornweighed 1,467g and apgar score was 4 and 8 at 1 and 5 min, intubated 5 min after birth due to bradycardia. Blood test showed anemia (Hb12.8g/dl) and cardiac ultrasonography showed myocardial hypertrophy and low cardiac wall motion. Histological analysis confirmed the diagnosis of placental chorioangioma. It is important that we should suspect a placental chorioangioma is one of the cause of fetal heart failure. O-058. DELTA-LIKE PROTEIN 1 AND NOTCH1 PROMOTER METHYLATION AND PROTEIN EXPRESSION IN PLACENTA Yota Shimanuki , Hiroyuki Mitomi , Shintaro Makino , Atsuo Itakura , Satoru Takeda , Takashi Yao . a Juntendo University Faculty of Medicine, Department of Obstetrics and Gynecology, Japan; b Juntendo University Faculty of Medicine, Department of Human Pathology, Japan; Dokkyo Medical University, Pathology, Japan Introduction: The establishment of a successful pregnancy requires the implantation of a competent blastocyst into a 'receptive' endometrium, facilitating the formation of a functional placenta. The Notch signalling family is a receptor-ligand family that regulates cellular processes as diverse as proliferation, apoptosis, differentiation, invasion and adhesion. Recently, some studies have identified Notch receptors and ligands in the endometrium, blastocyst and placenta. To investigate the methylation status of Delta-like protein 1 (Dll1) and Notch1 by methylation-specific PCR (MSP) and expression of those proteins using immunohistochemistry (IHC). Material and Methods: One hundred fifty patients, including early-onset PE (n1⁄413), late-onset PE (n1⁄425), FGR (n1⁄446), chorioamnionitis (n1⁄414), fetal anomaly (n1⁄425), and other disorders were recruited for this study. MSP and IHC were performed on formalin-fixed paraffin-embedded placental tissue blocks. Genomic DNA was extracted from 10mm sections