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Effect of PTCH1 expression on placental and fetal growth in rats treated with retinoic acid
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Abstracts / Placenta 59 (2017) 169e184
and IGF-I on proliferation and angiogenesis of trophoblast cells by using the human choriocarcinoma cell-line, BeWo. Materials & Methods: After incubating Bewo under the presence of TNF-a (10-105 pg/ml) and IGF-I (102 ng/mL), we assessed cell number by WST-1 assay, proliferation by BrdU uptake assay, and apoptosis by Caspase-3, 8 activity assay. We also measured the concentrations of VEGF, PlGF, and FGF in the conditioned media by ELISA . Results: Under the presence of IGF-I, cell number and BrdU uptake was dose-dependently enhanced by TNF-a (10-102pg/ml), while no such effects was detected without IGF-I. Caspase-3,8 activities were most suppressed in 102 pg/ml and most enhanced in 104-105 pg/ml of TNF-a. Caspase-3,8 activities were significantly suppressed by IGF-I regardless of TNF-a concentrations. The concentrations of PlGF and VEGF in the media were significantly high with 10-102 pg/ml of TNF-a, while TNF-a did not have any effects on FGF concentrations. Addition of IGF-I dose-dependently suppressed PlGF secretion and enhanced VEGF secretion. Conclusion: TNF-a and IGF-I influence on proliferation and angiogenesis of BeWo through independent and synergetic effects, which suggests that TNF-a and IGF-I are responsible for placental hypertrophy in pregnancy with obesity.
7. ONE FGR FETUS WITH PLACENTAL MESENCHYMAL DYSPLASIA IN DICHORIONIC DIAMNIOTIC TWIN PREGNANCY Shoko Jitsumori. Department of Obsterics and Gynecology, Wakayama Medical University, Japan Introduction: Placental mesenchymal dysplasia (PMD) is defined as swelling stem villi with hyperplasia and placentomegaly without trophoblast proliferation. We reported a case in which one FGR fetus showed PMD in a dichorionic-diamniotic (DD) twin pregnancy. Case: A 24-year-old woman, para 1, was referred to our hospital at 24 +1 weeks’ gestation due to one fetal growth restriction (FGR) (-2.6SD) and ipsilateral placental abnormality in DD twin. By ultrasound examination, the placenta of the FGR fetus had multiple cysts and was bulky along with its site. The other fetal placenta had no cyst. Maternal serum hCG level was 44,084 mIU/mL and not so high, which suspected placental mesenchymal dysplasia (PMD). The FGR fetus had no ultrasound findings of characteristics of Beckwith-Wiedemann syndrome. MRI at 32 weeks’ gestation also showed the presence of many small cysts in the placenta at the side of FGR fetus. As the growth of biparietal diameter of FGR fetus stopped (-4.5SD at 32 weeks’ gestation), a cesarean section was performed at 32+5 weeks’ gestation. The first neonate was a female, weighed 1799g (-0.1SD) with Apgar Score 7/8. The second one was a female, weighed 1215g (-2.5SD) with Apgar Score 8/9. The placenta weighed 1066g (above 90th percentile) at delivery and the chorionic vessels on the placental surface of the SGA birth were prominently enlarged. Macroscopically, the lesion of PMD occupied about 37% of the placenta. Pathological examination showed swelling stem villi without trophoblast proliferation, enlarged vessels and increase of interstitial cells. The immunostaining of p57kip2 was negative in the interstitial cells of swelling stem villi. In addition, the expression of p57kip2 was almost completely negative in trophoblasts of the abnormal villi and weakly positive in the trophoblast of normal-looking villi. Conclusion: It was estimated that extensive PMD occurred in one placenta of DD twin pregnancy, which lead to early-onset FGR.
9. EFFECT OF PTCH1 EXPRESSION ON PLACENTAL AND FETAL GROWTH IN RATS TREATED WITH RETINOIC ACID
model with low birth weight, we investigated an inhibitory effect on placental and fetal growth by PTCH1 expression due to RA administration to elucidate one of the causes of fetal growth restriction. Methods: Birth weight and crown-rump length (CRL) were compared between Sprague-Dawley rat (embryonic day 10) orally administered with RA 60 mg / kg (n ¼ 6) and non-administration group (n ¼ 14). In addition, gene expression levels of PTCH1 and Gli1 in placenta, heart, liver, intestinal tract, skin, brain were compared by using quantitative PCR. Results: Birth weight / CRL was significantly lower in the RA administration group (4.9 g / 3.9 cm) compared with the RA non-administration group (5.9 g / 4.6 cm). In neonatal rats treated with RA, myelomeningocele was present in 83.5%. The spinal cord tissue appeared more differentiated. Expression level of PTCH1 of the skin and intestinal tract increased in RAtreated rat, but the expression in the liver was low, and there was no difference in heart and brain. Expression level of Gli1 was increased in the skin and intestinal tract, with no significant difference in other tissues. Although expression level of PTCH1 in the placenta tended to be higher in the RA-treated group, there was no difference in expression level of Gli1. Conclusion: Fetal growth in rat fetus were inhibited by RA administration, but expression level of Gli1 did not decrease despite the increased expression level of PTCH1, suggesting a regulation of Gli1 expression by other than hedgehog pathway.
10. CONSERVATIVE MANAGEMENT OF A RETAINED PLACENTA: A REPORT OF FIVE CASES Mari Tomiie. Osaka Saiseikai Nakatsu Hosipital, Japan Objective: To clarify the risk factors for successful conservative treatment of retained placenta after vaginal delivery. Methods: Five women diagnosed as having placenta accreta between June 2013 and June 2015 were included. Gestational age, parity, history of abortion, uterine artery embolization (UAE), manual placenta removal, blood loss at delivery, retained placenta size, and outcome were evaluated. Results: In cases 1-4, retained placenta was detected on ultrasonography and magnetic resonance imaging (MRI) after manual placenta removal. UAE was performed for persistent bleeding in cases 2-5. As the hemorrhage was mild in cases1 and 5, a conservative approach was decided. Case 1: Nine months after conservative treatment, the retained placenta disappeared spontaneously. Case 2: Forty weeks after UAE, the retained placenta disappeared. Case 3: After UAE, no bleeding and decreased size of the mass in the uterus were observed. Case 4: The retained placenta was 10 cm in diameter. Persistent puerperal fever with leukocytosis and high C-reactive protein level developed during the postpartum period despite antibiotic treatment. The patient developed sepsis, and an urgent hysterectomy was performed on postpartum day 20. Case 5: After delivery, no signs of placenta detachment were observed. The entire placenta was left in situ after conservative treatment. The fever and leukocytosis subsided after antibiotic treatment: however, the patient developed a fever gradually. Hysterectomy was performed on postpartum day 13. Conclusion: Successful conservative management of retained placenta may affect the size of the retained placenta.
14. REGULATORY MECHANISM OF EXTRAVILLOUS TROPHOBLAST
CD59
EXPRESSION
ON
HUMAN
Masashi Ueda. Kyoto University Graduate School of Medicine, Japan Kazuhiro Kajiwara. Jikei University School of Medicine, Japan Objective: Pacthed-1 (PTCH1) suppresses hedgehog signaling. Decreased expression of PTCH1 induces Gli1, a downstream transcription factor, leading to cell proliferation. Retinoic acid (RA) has been reported to induce PTCH1 expression. Since RA administration induce myelomeningocele rat
Objective: In human placenta, endovascular extravillous trophoblast (eEVT) makes direct contact with complement components contained in the maternal blood. Although activated complement components could exert the cytotoxic effects, eEVT is not eliminated by their activation. We focused on possible avoidance mechanism inherent in eEVT.
Abstracts / Placenta 59 (2017) 169e184
and IGF-I on proliferation and angiogenesis of trophoblast cells by using the human choriocarcinoma cell-line, BeWo. Materials & Methods: After incubating Bewo under the presence of TNF-a (10-105 pg/ml) and IGF-I (102 ng/mL), we assessed cell number by WST-1 assay, proliferation by BrdU uptake assay, and apoptosis by Caspase-3, 8 activity assay. We also measured the concentrations of VEGF, PlGF, and FGF in the conditioned media by ELISA . Results: Under the presence of IGF-I, cell number and BrdU uptake was dose-dependently enhanced by TNF-a (10-102pg/ml), while no such effects was detected without IGF-I. Caspase-3,8 activities were most suppressed in 102 pg/ml and most enhanced in 104-105 pg/ml of TNF-a. Caspase-3,8 activities were significantly suppressed by IGF-I regardless of TNF-a concentrations. The concentrations of PlGF and VEGF in the media were significantly high with 10-102 pg/ml of TNF-a, while TNF-a did not have any effects on FGF concentrations. Addition of IGF-I dose-dependently suppressed PlGF secretion and enhanced VEGF secretion. Conclusion: TNF-a and IGF-I influence on proliferation and angiogenesis of BeWo through independent and synergetic effects, which suggests that TNF-a and IGF-I are responsible for placental hypertrophy in pregnancy with obesity.
7. ONE FGR FETUS WITH PLACENTAL MESENCHYMAL DYSPLASIA IN DICHORIONIC DIAMNIOTIC TWIN PREGNANCY Shoko Jitsumori. Department of Obsterics and Gynecology, Wakayama Medical University, Japan Introduction: Placental mesenchymal dysplasia (PMD) is defined as swelling stem villi with hyperplasia and placentomegaly without trophoblast proliferation. We reported a case in which one FGR fetus showed PMD in a dichorionic-diamniotic (DD) twin pregnancy. Case: A 24-year-old woman, para 1, was referred to our hospital at 24 +1 weeks’ gestation due to one fetal growth restriction (FGR) (-2.6SD) and ipsilateral placental abnormality in DD twin. By ultrasound examination, the placenta of the FGR fetus had multiple cysts and was bulky along with its site. The other fetal placenta had no cyst. Maternal serum hCG level was 44,084 mIU/mL and not so high, which suspected placental mesenchymal dysplasia (PMD). The FGR fetus had no ultrasound findings of characteristics of Beckwith-Wiedemann syndrome. MRI at 32 weeks’ gestation also showed the presence of many small cysts in the placenta at the side of FGR fetus. As the growth of biparietal diameter of FGR fetus stopped (-4.5SD at 32 weeks’ gestation), a cesarean section was performed at 32+5 weeks’ gestation. The first neonate was a female, weighed 1799g (-0.1SD) with Apgar Score 7/8. The second one was a female, weighed 1215g (-2.5SD) with Apgar Score 8/9. The placenta weighed 1066g (above 90th percentile) at delivery and the chorionic vessels on the placental surface of the SGA birth were prominently enlarged. Macroscopically, the lesion of PMD occupied about 37% of the placenta. Pathological examination showed swelling stem villi without trophoblast proliferation, enlarged vessels and increase of interstitial cells. The immunostaining of p57kip2 was negative in the interstitial cells of swelling stem villi. In addition, the expression of p57kip2 was almost completely negative in trophoblasts of the abnormal villi and weakly positive in the trophoblast of normal-looking villi. Conclusion: It was estimated that extensive PMD occurred in one placenta of DD twin pregnancy, which lead to early-onset FGR.
9. EFFECT OF PTCH1 EXPRESSION ON PLACENTAL AND FETAL GROWTH IN RATS TREATED WITH RETINOIC ACID
model with low birth weight, we investigated an inhibitory effect on placental and fetal growth by PTCH1 expression due to RA administration to elucidate one of the causes of fetal growth restriction. Methods: Birth weight and crown-rump length (CRL) were compared between Sprague-Dawley rat (embryonic day 10) orally administered with RA 60 mg / kg (n ¼ 6) and non-administration group (n ¼ 14). In addition, gene expression levels of PTCH1 and Gli1 in placenta, heart, liver, intestinal tract, skin, brain were compared by using quantitative PCR. Results: Birth weight / CRL was significantly lower in the RA administration group (4.9 g / 3.9 cm) compared with the RA non-administration group (5.9 g / 4.6 cm). In neonatal rats treated with RA, myelomeningocele was present in 83.5%. The spinal cord tissue appeared more differentiated. Expression level of PTCH1 of the skin and intestinal tract increased in RAtreated rat, but the expression in the liver was low, and there was no difference in heart and brain. Expression level of Gli1 was increased in the skin and intestinal tract, with no significant difference in other tissues. Although expression level of PTCH1 in the placenta tended to be higher in the RA-treated group, there was no difference in expression level of Gli1. Conclusion: Fetal growth in rat fetus were inhibited by RA administration, but expression level of Gli1 did not decrease despite the increased expression level of PTCH1, suggesting a regulation of Gli1 expression by other than hedgehog pathway.
10. CONSERVATIVE MANAGEMENT OF A RETAINED PLACENTA: A REPORT OF FIVE CASES Mari Tomiie. Osaka Saiseikai Nakatsu Hosipital, Japan Objective: To clarify the risk factors for successful conservative treatment of retained placenta after vaginal delivery. Methods: Five women diagnosed as having placenta accreta between June 2013 and June 2015 were included. Gestational age, parity, history of abortion, uterine artery embolization (UAE), manual placenta removal, blood loss at delivery, retained placenta size, and outcome were evaluated. Results: In cases 1-4, retained placenta was detected on ultrasonography and magnetic resonance imaging (MRI) after manual placenta removal. UAE was performed for persistent bleeding in cases 2-5. As the hemorrhage was mild in cases1 and 5, a conservative approach was decided. Case 1: Nine months after conservative treatment, the retained placenta disappeared spontaneously. Case 2: Forty weeks after UAE, the retained placenta disappeared. Case 3: After UAE, no bleeding and decreased size of the mass in the uterus were observed. Case 4: The retained placenta was 10 cm in diameter. Persistent puerperal fever with leukocytosis and high C-reactive protein level developed during the postpartum period despite antibiotic treatment. The patient developed sepsis, and an urgent hysterectomy was performed on postpartum day 20. Case 5: After delivery, no signs of placenta detachment were observed. The entire placenta was left in situ after conservative treatment. The fever and leukocytosis subsided after antibiotic treatment: however, the patient developed a fever gradually. Hysterectomy was performed on postpartum day 13. Conclusion: Successful conservative management of retained placenta may affect the size of the retained placenta.
14. REGULATORY MECHANISM OF EXTRAVILLOUS TROPHOBLAST
CD59
EXPRESSION
ON
HUMAN
Masashi Ueda. Kyoto University Graduate School of Medicine, Japan Kazuhiro Kajiwara. Jikei University School of Medicine, Japan Objective: Pacthed-1 (PTCH1) suppresses hedgehog signaling. Decreased expression of PTCH1 induces Gli1, a downstream transcription factor, leading to cell proliferation. Retinoic acid (RA) has been reported to induce PTCH1 expression. Since RA administration induce myelomeningocele rat
Objective: In human placenta, endovascular extravillous trophoblast (eEVT) makes direct contact with complement components contained in the maternal blood. Although activated complement components could exert the cytotoxic effects, eEVT is not eliminated by their activation. We focused on possible avoidance mechanism inherent in eEVT.