Downregulation of PGRMC1 accelerates differentiation and fusion of a human trophoblast cell line.

Mononuclear cytotrophoblasts (CTs) differentiate and fuse to form multinuclear syncytiotrophoblasts (STs), which produce human chorionic gonadotropin (hCG) and progesterone to maintain pregnancy. Impaired differentiation and fusion of CTs to form STs are associated with hypertensive disorders of pregnancy and fetal growth restriction. Progesterone receptor membrane component 1 (PGRMC1) is a multifunctional single transmembrane heme-binding protein. We previously demonstrated that downregulation of PGRMC1 promotes endometrial stromal cell differentiation (decidualization). Here, we explored the role of PGRMC1 in trophoblast differentiation and fusion. PGRMC1 expression was lower in STs than in CTs of first-trimester placental tissues. PGRMC1 expression in BeWo cells (a trophoblast-derived choriocarcinoma cell line) decreased upon dibutyryl-cAMP (db-cAMP)-induced differentiation. Both inhibition and knockdown of PGRMC1 stimulated hCG production in the presence of db-cAMP. Furthermore, a quantitative cell fusion assay we developed revealed that inhibition and knockdown of PGRMC1 enhanced db-cAMP-stimulated cell fusion. Peroxisome proliferator-activated receptor γ (PPARγ) agonists decreased PGRMC1 expression and stimulated the cell fusion in BeWo cells. These findings suggest that downregulation of PGRMC1 expression in part through activation of PPARγ during trophoblast differentiation promotes hCG production and cell fusion for formation and maintenance of placental villi during pregnancy.

Alpha-1 Antitrypsin-Induced Endoplasmic Reticulum Stress Promotes Invasion by Extravillous Trophoblasts.

Alpha-1 antitrypsin (A1AT) is a glycoprotein that has been shown to protect tissues from proteolytic damage under various inflammatory conditions. Several studies show that A1AT may be associated with pre-eclampsia. However, the role of A1AT expression in placental physiology is not fully understood. In the present study, we aim to characterize the expression and function of placental A1AT. A1AT knockdown is found to reduce the expression of the serine protease HTRA1 in a trophoblast cell line. In addition, A1AT overexpression (A1AT-OE) increases the expression of HTRA1, IL6, CXCL8, and several markers of endoplasmic reticulum (ER) stress. Treatment with tunicamycin or thapsigargin, which induces ER stress, increases HTRA1 expression. Furthermore, immunohistochemistry reveals that HTRA1 is expressed in trophoblasts and the endometrial decidual cells of human placentas. An invasion assay shows that A1AT and HTRA1 stimulate cell invasion, but treatment with the ER stress inhibitors reduces the expression of HTRA1 and ER stress markers and prevents cell invasion in A1AT-OE trophoblasts. These results suggest that endogenous A1AT regulates inflammatory cytokine expression and HTRA1-induced trophoblast invasion via the induction of ER stress. It is concluded that an imbalance in the functional link between A1AT and ER stress at the maternal-fetal interface might cause abnormal placental development.

Complications and reproductive outcome after uterine artery embolization for retained products of conception.

AIM: The purpose of this study was to evaluate the complications and reproductive outcome after uterine artery embolization (UAE) for retained products of conception (RPOC). METHODS: This was a retrospective medical-records review study of 57 women treated for RPOC. Participants were divided into two groups: women who underwent treatment with UAE (UAE group: n = 32, 56.1%) and those without UAE (control group: n = 25, 43.9%). The complications and reproductive outcomes were compared between the two groups. Information on subsequent pregnancies and their outcomes was available for 30 women who attempted to conceive. RESULTS: There were no significant differences in the interval from the last delivery or abortion (40.1 ± 3.4 vs 51.0 ± 5.1 months, respectively; P = 0.16), the rate of severe bleeding under hysteroscopy (18.5 vs 9.1%, respectively; P = 0.65), the conception rate (58.8 vs 61.5%, respectively; P = 1.0) and mean time to conception (9.9 ± 1.6 vs 11.0 ± 2.9 months, respectively; P = 0.17) in women in the UAE group compared with those in the control group. Rates of post-partum hemorrhage (PPH) and manual removal of placenta (25.0% in the UAE group and 16.7% in the control group, respectively) were higher than the general population. CONCLUSION: Selective UAE for RPOC may be a preferable procedure in women who are suspected as having a risk of severe bleeding under treatment. Women who were treated for RPOC, regardless of UAE, were at risk of PPH and difficulty in removing the placenta in future pregnancies.

Perineal pyoderma gangrenosum in pregnancy: A case report.

Pyoderma gangrenosum is a rare ulcerating neutrophilic dermatosis. We describe the case of a 28-year-old woman with pyoderma gangrenosum in the perineal region during pregnancy. Cytological analysis of a skin biopsy specimen showed neutrophilic infiltrates across all the layers of the dermis, confirming the diagnosis of pyoderma gangrenosum. Determining a management plan, including the mode of delivery, was difficult. Oral prednisolone was started and her ulcer started to improve, but she still had the ulcer when she reached full term. Because there was a concern that the ulcer would be worsened by vaginal delivery, cesarean section was performed. After her delivery, pyoderma gangrenosum had not appeared at the cesarean incision and the ulcer in the perineal region had improved. Obstetricians should be aware of pyoderma gangrenosum as a differential diagnosis when vulvar ulceration develops during pregnancy.

Placental Elasticity as a New Non-invasive Predictive Marker of Pre-eclampsia.

Point shear wave elastography is an ultrasonography technique used to evaluate tissue elasticity. We examined whether placental elasticity is useful for predicting the onset of pre-eclampsia. Two hundred twenty-one participants were divided into two groups: one group at low risk (n = 185) and the other at high risk (n = 36) for pre-eclampsia. The two groups were compared with respect to shear wave velocity (SWV) of the placenta. Use of SWV as a predictor of pre-eclampsia was also investigated by creating a receiver operating characteristic (ROC) curve. The ROC curve was used to set a cutoff SWV value for predicting pre-eclampsia. The SWV of the high-risk group was significantly higher than that of the low-risk group (p < 0.001). Thirteen participants developed pre-eclampsia after SWV measurements, and the SWVs of these participants were significantly higher than those of participants in who pre-eclampsia did not develop. The cutoff value and area under the ROC curve were 1.188 m/s and 0.9118, respectively. Placental elasticity was significantly increased even before the onset of pre-eclampsia onset and, thus, may be a parameter used to predict the onset of pre-eclampsia.

Serum soluble LIGHT in the early third trimester as a novel biomarker for predicting late-onset preeclampsia.

Our aim was to evaluate whether serum levels of soluble LIGHT (sLIGHT) at 27-31 weeks can predict the later occurrence of gestational hypertension (GH), late-onset preeclampsia (PE), and early-onset PE. Mean blood pressure (MBP), soluble fma-like tyrosine kinase 1/placental growth factor (sFlt-1/PlGF) ratio at 27-31 weeks, and sLIGHT at 27-31 weeks were independent risk factors for late-onset PE. The combination of the three risk factors improved sensitivity with a false-positive rate of 10% (MBP: 60%, log10(sFlt-1/PlGF): 45%, sLIGHT: 35%, combination: 75%). Serum sLIGHT in the early third trimester may be a novel biomarker for predicting late-onset PE.

Fibrosis in Preeclamptic Placentas Is Associated with Stromal Fibroblasts Activated by the Transforming Growth Factor-ß1 Signaling Pathway.

Although fibrosis is one of the most prominent pathologic features of preeclamptic (PE) placentas, its mechanism remains largely unknown. Consistent with previous reports, we observed overexpression of collagen; actin, α2, smooth muscle, aorta; connective tissue growth factor; and fibronectin in PE placentas compared with control ones. To investigate the mechanism of fibrosis in PE placentas, placental fibroblasts were isolated from PE placentas or normal pregnancies at delivery. The expression of fibrosis-related factors in fibroblasts was evaluated by real-time RT-PCR, Western blotting, enzyme-linked immunosorbent assay, and gene microarrays. An in vitro collagen gel contraction assay was also performed. Fibroblasts isolated from PE placentas showed higher expression levels of fibrosis-related factors compared with those from control ones. Global gene expression profiling of PE fibroblasts was contrasted with that of control ones and indicated an intimate association with transforming growth factor-ß1 (TGFB1) signaling. Furthermore, the PE fibroblasts expressed abundant phosphorylated SMAD family member 2 and showed higher expression levels of target genes of TGFB1 signaling compared with the control ones. The PE fibroblasts also had a greater ability to contract compared with the control ones. Contractility also depended on TGFB1 signaling. Our results suggest that TGFB1 signaling is activated in the fibroblasts in PE placentas and that these active fibroblasts contribute to fibrosis.

Vascular endothelial growth factor receptor tyrosine kinase inhibitor PTK787/ZK 222584 inhibits both the induction and elicitation phases of contact hypersensitivity.

Vascular endothelial growth factor (VEGF) and its endothelial cell receptors (VEGFR) have been shown to be involved in the pathogenesis of the contact hypersensitivity (CHS) reaction. Previous studies have demonstrated that anti-VEGFR-2 antibody significantly suppresses the elicitation phase of CHS but does not affect the induction phase. PTK787/ZK 222584 (1-[4-chloroanilino]-4-[4-pyridylmethyl] phthalazine succinate; PTK/ZK) is a potent inhibitor of VEGFR tyrosine kinases. To test the effect of PTK/ZK on the induction and elicitation phases of CHS separately, we used an established method of CHS assay-sensitization and challenge in BALB/c mice. Either 50 mg/kg/day PTK/ZK or vehicle serving as a control was administered orally in the induction or elicitation phases separately. In the afferent phase, flow cytometry of skin-draining lymph node cells revealed that the migration of Langerhans cells was suppressed in the mice treated with PTK/ZK at sensitization. The degrees of ear swelling at 24 and 48 h were significantly diminished in mice treated with PTK/ZK at sensitization (P < 0.05). In the efferent phase, the degrees of ear swelling at 24 h (P < 0.01) and 48 h (P < 0.05), ear blood flow at 24 and 48 h (P < 0.01), and production of VEGF in the epidermis at 24 h (P < 0.05) were significantly suppressed in mice treated with PTK/ZK at elicitation. These findings and previous demonstrations suggest that both VEGF R-1 and VEGF R-2 are needed during the induction phase, and that VEGFR-2 has a pivotal role in the elicitation phase of the CHS reaction.

[The effect of Sairei-to on oxygen-induced retinal neovascularization in the neonatal rat].

PURPOSE: To study the effect of Sairei-to (ST), a Japanese traditional medicine, on oxygen-induced retinopathy (OIR) in rats. METHODS: OIR was induced by maintaining Sprague-Dawley neonatal rats in 80% oxygen for 12 days. The rats were treated once daily with oral administration of 0.75 g/kg (n = 9), 1.5 g/kg (n = 13) of ST in water, or water alone (WA, n = 13) at 5 mL/ kg body weight from day 6 to day 17. On day 18, retinal samples were collected. Retinal neovascularization (NV) was assessed by the NV score, and by the percentage of avascular area (% AVA), using a method previously reported. The number of severe retinal NV cases (NV > or = 9) was compared. The retinal vascular endothelial growth factor (VEGF) concentrations were measured with an immunoassay kit, at 0, 12, 24, 72 and 144 hours after oxygenation. RESULTS: NV score and % AVA decreased in the ST treated group compared to the WA group. However, severe NV was seen in five cases of WA and in one case of the ST treated group. Thus severe NV was inhibited significantly by ST treatment (p = 0.0185). Retinal VEGF did not differ between groups at any time points. CONCLUSION: These data suggest that severe NV in OIR is inhibited by ST treatment.