The Expression and Significance of Matrix Metalloproteinase, Wnt5a and β-Catenin in Placental Tissue of Preeclampsia

In order to investigate the expression and clinical significance of MMPs (Matrix Metalloproteinase) (MPP-2 and MPP-9), Wnt5a as well as β-catenin in placental tissues of preeclampsia (PE) patients, 36 PE patients (PE group) and 25 pregnant women (control group) with normal pregnancy were selected for cesarean delivery. The expression and localization of MPP-2, MPP-9, Wnt5a as well as β-catenin proteins in placental tissue were detected by Western blot method and immunohistochemistry method. The correlation between the poor prognosis of mother and child and the expression level of MMPs, Wnt5a as well as β-catenin is analyzed. The results showed that Western blot results showed that MPP-2, MPP-9, Wnt5a as well as β-catenin proteins were expressed in placental tissue, and the expression level of them of placental tissue in PE group was significantly lower than that in control group (P < 0.01). Immunohistochemistry showed that MMPs, Wnt5a, as well as β-catenin proteins were expressed in the cytotrophoblast cell (CTB) and syncytiotrophoblast cell (STB) of placental tissue, among which Wnt5a proteins as well as β-catenin proteins were mainly expressed in the cytoplasm of STB and CTB respectively. MMPs proteins were mainly expressed in the cytoplasm of these two types of cells. Low expression of MPPs, Wnt5a, β-catenin proteins may be related to blood pressure as well as 24 h urine protein of pregnant women at admission. In conclusion, Low expression of MPPs, Wnt5a, β-catenin proteins may be involved in the PE pathogenesis.

Human Cytomegalovirus modifies placental small extracellular vesicle secretion and composition towards a proviral phenotype to enhance infection of fetal recipient cells

Although placental small extracellular vesicles (sEVs) are extensively studied in the context of pregnancy, little is known about their role during human cytomegalovirus (hCMV) congenital infection, especially at the beginning of pregnancy. In this study, we examined the consequences of hCMV infection on sEVs production and composition using an immortalized human cytotrophoblast cell line derived from first trimester placenta. By combining complementary approaches of biochemistry, imaging techniques and quantitative proteomic analysis, we showed that hCMV infection increased the yield of sEVs produced by cytotrophoblasts and modified their protein composition towards a proviral phenotype. We further demonstrated that sEVs secreted by hCMV-infected cytotrophoblasts potentiated infection in naive recipient cells of fetal origin, including neural stem cells. Importantly, the enhancement of hCMV infection was also observed with sEVs prepared from either an ex vivo model of infected histocultures from early placenta or from the amniotic fluid of patients naturally infected by hCMV at the beginning of pregnancy. Based on these findings, we propose that placental sEVs could be key actors favoring viral dissemination to the fetal brain during hCMV congenital infection.

Association of sflt-1 as a maternal serum biomarker in preeclampsia: A case–control tertiary care hospital based study

Objectives: Preeclampsia (PE) is a multisystemic disorder portrayed by the new beginning of circulatory pressure more noteworthy than 140/90 mmHg and proteinuria with 0.3 g in a 24 h on dip stick emerging after 20 weeks of incubation. The hidden pathophysiology of PE includes endothelial brokenness and vasospasm beginning principally in the placenta. The unusual growth of blood vessels in placenta leads to poor perfusion. This relative hypoxic condition in placenta causes arrival of antiangiogenic factors into the maternal blood dissemination which prompts the modifications in maternal fundamental endothelial functions and causes hypertension. Soluble fms-like tyrosine kinase (sFlt) can form a heterodimer, binding with vascular endothelial growth Factor A and placental growth factor. In preeclamptic subjects, there will be an imbalance in anti-angiogenesis factors and there will be incomplete arterial transformation and cytotrophoblast cell division. Due to imbalance in sFlt levels in preeclamptic women it effects in the blood vessels by constriction and leads to endothelial dysfunction. This study aim is to compare the maternal serum concentration of sFlt levels in normotensive pregnant women to preeclamptic women in early and late gestational weeks. Material and Methods: Out of 300 participants in the case–control study, 150 were preeclamptic women as cases and 150 as normotensive pregnant women as controls participated in the present study. A 5 ml of maternal venous blood was collected; the serum was separated and stored at –800°C till the analysis. Using commercially available enzyme-linked immunosorbent assay (ELISA) kits from Chongqing Biospes Co., Ltd., (suppliers: Infobio Company, New Delhi) was measured with ELISA microplate reader at 450 nm (Merilyzer Eiaquant Company). Results: Out of 300 participants in the study, 46 pregnant women were early gestational weeks and 254 were late gestational weeks. The complications due to severe PE such as intrauterine death are 15%, intrauterine fetal growth retardation 33%, and premature 15%. The statistical analyses were performed by Statistical Packages for the Social Sciences Software 22. The area under the receiver operating characteristic curve is 0.82, with 91% sensitivity, and 79% specificity. The significance in the maternal serum sFlt levels was calculated by the Mann–Whitney U-test. By comparing the cases and controls, it was found that maternal serum sFlt1 were significantly higher in preeclamptic women with Z = 2.96 and U = 9021 with P = 0.005 significance. Conclusion: This is the first South Indian study. If we compare the sFlt1 levels in early and late gestational weeks, in late gestational weeks in controls and PE the levels were highly significant than early gestational weeks of PE and controls. Maternal serum sFlt can be used as a preeclamptic diagnostic marker in South Eastern Kolar population.

The Cannabinoid Delta-9-tetrahydrocannabinol Disrupts Estrogen Signaling in Human Placenta

Cannabis consumption is increasing worldwide either for recreational or medical purposes. Its use during gestation is associated with negative pregnancy outcomes such as, intrauterine growth restriction, preterm birth, low birth weight, and increased risk of miscarriage, though the underlying molecular mechanisms are unknown. Cannabis sativa main psychoactive compound, Δ9-tetrahydrocannabinol (THC) is highly lipophilic, and as such, readily crosses the placenta. Consequently, THC may alter normal placental development and function. Here, we hypothesize alterations of placental steroidogenesis caused by THC exposure. The impact on placental estrogenic signaling was examined by studying THC effects upon the enzyme involved in estrogens production, aromatase and on estrogen receptor α (ERα), using placental explants, and the cytotrophoblast cell model BeWo. Aromatase expression was upregulated by THC, being this effect potentiated by estradiol. THC also increased ERα expression. Actions on aromatase were ERα-mediated, as were abolished by the selective ER downregulator ICI-182780 and dependent on the cannabinoid receptor CB1 activation. Furthermore, the presence of the aromatase inhibitor Exemestane did not affect THC-induced increase in ERα expression. However, THC effects on ERα levels were reversed by the antagonists of CB1 and CB2 receptors AM281 and AM630, respectively. Thus, we demonstrate major alterations in estrogen signaling caused by THC, providing new insight on how cannabis consumption leads to negative pregnancy outcomes, likely through placental endocrine alterations. Data presented in this study, together with our recently reported evidence on THC disruption of placental endocannabinoid homeostasis, represent a step forward into a deeper comprehension of the puzzling actions of THC.

MEF2 transcription factors in human placenta and involvement in cytotrophoblast invasion and differentiation

Development of the human placenta and its trophoblast cell types is critical for a successful pregnancy. Defects in trophoblast invasion and differentiation are associated with adverse pregnancy outcomes, including preeclampsia. The members of myocyte enhancer factor-2 (MEF2) family of transcription factors are key regulators of cellular proliferation, differentiation, and invasion in various cell types and tissues and might play a similarly important role in regulating trophoblast proliferation, invasion, and differentiation during human placental development. In the present study, using human cytotrophoblast cell lines (HTR8/SVneo and BeWo) and primary human cytotrophoblasts (CTBs), we show that members of the MEF2 family are differentially expressed in human placental CTBs, with MEF2B and MEF2D being highly expressed in first trimester extravillous CTBs. Overexpression of MEF2D results in cytotrophoblast proliferation and enhances the invasion and migration of extravillous-like HTR8/SVneo cells. This invasive property is blocked by overexpression of a dominant negative MEF2 (dnMEF2). In contrast, MEF2A is the principal MEF2 isoform expressed in term CTBs, MEF2C and MEF2D being expressed more weakly, and MEF2B expression being undetected. Overexpression of MEF2A induces cytotrophoblast differentiation and syncytium formation in BeWo cells. During in vitro differentiation of primary CTBs, MEF2A expression is associated with CTB differentiation into syncytiotrophoblast. Additionally, the course of p38 MAPK and ERK5 activities parallels the increase in MEF2A expression. These findings suggest individual members of MEF2 family distinctively regulate cytotrophoblast proliferation, invasion, and differentiation. Dysregulation of expression of MEF2 family or of their upstream signaling pathways may be associated with placenta-related pregnancy disorders.

IMMUNOHISTOCHEMICAL STUDY OF METALLOPROTEINASES-2 IN THE TROPHOBLAST IN CASE OF TORCH-INFECTIONS ON THE MATERIAL OF ABORTIONS AT THE GESTATIONAL TERM OF 5-6 WEEKS

This study is a fragment of a series of immunohistochemical studies of trophoblast in case of TORCH-infections, which are scheduled at different times of gestation. This article focuses on the results of studies of the trophoblast in gestational age 5-6 weeks. The aim of the study was by means of the immunohistochemical method to determine the features of expression of MP-2 in different types of trophoblast with TORCH-infections as compared to observations without an infectious process. Abortion material of 5-6 weeks of gestation was investigated. The main group included 16 observations of TORCH-infections, and the control - 14 observations of interrupted pregnancies with no signs of infection (abortion for social reasons). Immunohistochemical method was applied on metalloproteinases-2 primary antibody and polymer system for visualization of antigen using diaminobenzidine manufactured by DAKO. Using computer microdensitometry in a specialized computer program ImageJ the optical density of color was assessed. According to the results of immunohistochemical studies using the method of microdensitometry at the gestational age of 5-6 weeks, as in TORCH-infections and infectious process, the greatest expression of metalloproteinase-2 is noted in invasive trophoblast, the lowest - in the synthitiotrophoblast of chorial villi, and intermediate rates are seen in cytotrophoblast chorial villi and cell columns. With TORCH-infections, the expression of metalloproteinase-2 is reduced in all four types of trophoblast (cytotrophoblast chorial villi; cytotrophoblast cell columns; invasive cytotrophoblast in endometrial fragments), with the exception of synthiotrophoblast of chorial villi.