Preeclampsia Decidua Research Articles

Altered 5-methylcytosine modification of mRNA is involved in the pathogenesis of pre-eclampsia.

5-Methylcytosine (m5 C) is a prevalent RNA modification in messenger RNAs (mRNAs). Despite its abundance, its role in the decidua of pre-eclampsia (PE)remains elusive. In this study, we utilized methylated RNA immunoprecipitation sequencing (MeRIP-seq)and RNA-sequencing (RNA-seq) to map m5 C peaks and mRNA expression profile in the decidua of human early-onset PE (EPE), late-onset PE (LPE), and normal pregnancy (NP). Gene Ontology and Kyoto Encyclopedia of Genes and Genomespathway analyses elucidated potential roles of the differentially methylated mRNAs (DMGs) and differentially expressed mRNAs in decidualization pathways. Integrative analysis of MeRIP-seq and RNA-seq data pinpointed 50 candidate genes linked to PE, marked by both differentially methylated m5 C peaks and congruent expression changes. To validate these observations, we selected nine genes for verification via quantitative PCR. Our results underscore the precision and reproducibility of our bioinformatics approach. Importantly, we propose that changes in m5 C modification and expression of relevant mRNA might influence the pathogenesis of PE by hampering decidualization. This work shines light on the distinct mRNA m5 C modification patterns and expression profiles in the decidua of PE, implicating pivotal signaling disruptions and decidualization impediments in the onset of PE.

Regulatory Effect of Vitamin D on Renin Expression at Maternal-Fetal Interface

To investigate the regulatory effect and mechanism of vitamin D on the local renin-angiotensin system at maternal-fetal interface in the pathological process of preeclampsia (PE). The mRNA and protein expression of renin in decidua of normal pregnancy and PE placentas was determined by RT-PCR and Western blot. Normal decidual tissues were treated with active and inactive vitamin D for 48 h in vitro and the expressions of renin and vitamin D deactivating enzyme CYP24A1 were determined by RT-PCR and Western blot. Normal decidual stromal cells and glandular epithelial cells were isolated and purified, and identified by immunocytochemical staining. RT-PCR was used to examine the mRNA of vdr, cyp27 b1, cyp24 a1, and renin in the two types of cells and in decidual tissue, and the mRNA products were subjected to gel electrophoresis. These two cell types were treated with active and inactive vitamin D in vitro and the expressions of renin and vitamin D deactivating enzyme CYP24A1 were determined by RT-PCR and Western blot. Decidual gland epithelial cells were treated with protein kinase A (PKA) activator forskolin or inhibitor H89 to explore the interaction between PKA pathway and vitamin D in the regulation of renin expression. The expression of renin in PE decidua was significantly higher than that of normal control at transcriptional and translational levels ( P<0.05). Vitamin D treatment could significantly down-regulate the expression of renin in normal decidua tissues ( P<0.05), while it significantly up-regulated CYP24A1 expression ( P<0.001). Decidual stromal cells and gland epithelial cells were successfully isolated from decidual tissue. Compared with that in decidual stromal cells, the mRNA level of vitamin D-related molecules in gland epithelial cells was more similar to that in decidual tissue. Active or inactive vitamin D treatment significantly inhibited the expression of renin in glandular epithelial cells ( P<0.05), but the expression of renin in decidual stromal cells was not affected. However, the treatment of active or inactive vitamin D in these two kinds of cells significantly increased the expression of CYP24A1 ( P<0.001). Active vitamin D could significantly inhibit the upregulation of renin by PKA agonist forskolin, and could inhibit the expression of renin through synergy with PKA inhibitor H89. The expression of renin in placental decidua is up-regulated in patients with PE, and the activation of local renin-angiotensin system at the maternal-fetal interface may be involved in the pathogenesis of PE. Vitamin D can specifically down-regulate renin expression in human decidual gland epithelial cells by competing with the PKA pathway. Vitamin D supplementation may have potential value for clinical intervention of PE.

Narrative review of the relationship between the maternal-fetal interface immune tolerance and the onset of preeclampsia.

Background and ObjectiveThe establishment of maternal-fetal interface immune tolerance is essential for successful pregnancy. Studies have shown that spontaneous abortion, recurrent abortion, and fetal growth restriction are related to maternal-fetal interface immune dysfunction. Preeclampsia is an idiopathic condition related to pregnancy which manifests as hypertension, proteinuria, and other target organ damage after 20 weeks of gestation. Although the etiology of preeclampsia is still unknown, its pathogenesis is thought to be related to genetics, environment, and metabolism. In recent years, more and more studies have been conducted on the mechanism of immune tolerance at the maternal-fetal interface and the relationship between immune dysregulation and the pathogenesis of preeclampsia. This paper summarizes the latest studies on this topic in order to find new insights into the pathogenesis of preeclampsia and make a reflection on clinical diagnosis and treatment in different phenotype of preeclampsia.MethodsThe research and latest progress published from 2000 to December 2021 on the relationship between maternal-fetal interface immune tolerance and preeclampsia were broadly retrieved and researched using PubMed and Web of Science databases.Key Content and FindingsThe mechanism of natural killer cells (NK cells) and macrophages at the maternal-fetal interface in immune tolerance, as well as their cytotoxicity and cytokine secretion dysfunction, may be related to the pathogenesis of preeclampsia. The expression of nonclassical type I human leukocyte antigen (HLA) on extravillous trophoblast (EVT) cell were down-regulated in decidua of preeclampsia, which may induce increase EVT death caused by activating of cytotoxic NK cell. In addition, genetic polymorphism of nonclassical type I HLA on the EVT cell membrane may be related to the pathogenesis of preeclampsia, although this is likely to be a combination of 3 nonclassical type I HLA genotypes and requires sequencing to verify.ConclusionsWe demonstrated how the maternal-fetal interface immune dysfunction contribute to the pathogenesis of preeclampsia, further study and clinical trial based on this theory of pathogenesis may reveal new immune treatment method of preeclampsia.

P-033. Changes of natural cytotoxicity receptors of CD56dim and CD56bright NK cells in peripheral blood and decidua in preeclampsia

P-033. Changes of natural cytotoxicity receptors of CD56dim and CD56bright NK cells in peripheral blood and decidua in preeclampsia

Pro-inflammatory mediators and signaling proteins in the decidua of pre-eclampsia.

To evaluate the role of CD68+ macrophages and inflammatory/signaling proteins in the decidua of singleton pregnancies with late-onset pre-eclampsia. This study was designed as a prospective case-control study. Decidual tissue samples were obtained from twenty healthy pregnant women as a control group and twenty pregnant women with late-onset pre-eclampsia showing severe symptoms as the study group. We examined the abundance of CD68+ macrophages in both groups using flow cytometry. Protein and mRNA expression levels of inflammatory/signaling proteins, including inducible nitric oxide synthase, nuclear factor-κB inhibitor α, cyclooxygenase-2, and phosphorylated c-Jun N-terminal kinase, in the decidua of both groups were measured using Western blotting and Reverse Transcription-Polymerase Chain Reaction, respectively. Student's t-tests were performed for statistical analysis. The numbers of CD68+ macrophages were similar in the study and control groups (p=0.47). However, the levels of inducible nitric oxide synthase, nuclear factor-κB, cyclooxygenase-2, and phosphorylated c-Jun N-terminal kinase were significantly increased in the study group. Therefore, pro-inflammatory mediators and signaling proteins in the decidua during pre-eclampsia may be related to the pathogenesis of pre-eclampsia. Pre-eclampsia-induced alterations in the expression of inflammatory/signaling proteins in the decidua during singleton pregnancies may play a critical role in the pathogenesis of pre-eclampsia.

T lymphocytes in the third trimester decidua in preeclampsia

ABSTRACTObjective: The aim of study was to conduct immunohistochemical quantification of CD3+ and CD8+ decidual lymphocytes in preeclampsia. Methods: A study group included 30 cases of preeclampsia and a control group included 20 healthy pregnant women, all delivered by Cesarean section. Samples of placental bed were analyzed after immunohistochemical staining of CD45+, CD3+ and CD8+ cells. Results: The group with preeclampsia included a significantly higher number of CD3+ (p < 0.01) and CD8+ (p < 0.05) T lymphocytes. Conclusion: It is certain that thebalance dysregulation of T cell of the immune milieu of deciduais of importance in etiopathogenesis and manifestations of preeclampsia.

169. Retinoic acid down regulates sFlt1 in decidual stromal cells independent of decidualization. Implications to preeclampsia

Introduction Retinoic acid (RA) plays a vital role during pre- and early pregnancy stages at the maternal-fetal interface (decidua). We recently showed that RA levels are reduced in the decidua of women with preeclampsia (PE). This finding, combined with the knowledge that PE decidual stromal cells (DSC) secrete elevated levels of the anti-angiogenic factor sFlt1 and that the promoter of this protein has a putative RA binding site, suggests the possibility that RA plays a direct role in regulating sFlt1 expression in DSC. This study was to determine the effects of RA on sFlt1 production in DSC. The relationship of these effects to DSC decidualization, known to be defective in PE, was also investigated. Methods Primary DSCs (n = 6 in each group, normotensive (NT) and PE) were treated with 0.1 μ M of RA, 1 μ M BMS493 (pan RA antagonist) or 0.5 mM cAMP (to induce decidualization) for 12 days. sFLT1 and PRL levels were determined by using both qPCR and ELISA. Group means were analyzed by Mann–Whitney U test. p ⩽ 0.05 is considered significant. Results RA treatment of NT-DSCs and PE-DSCs inhibited sFlt1 production to 44 ± 24% and 24 ± 13% of controls but did not affect DSC decidualization as quantified by prolactin (PRL) secretion. Independent of RA, cAMP-mediated decidualization reduced sFlt1 and increased PRL levels in NT-DSC but only moderately in PE-DSC. BMS493 blocked the effects of RA and, by itself, induced upregulation of sFlt1. Studies using decidualization defective stromal cells confirmed that reduction of sFlt1 by RA is independent of decidualization. Conclusion RA downregulates sFlt1 production in DSCs independent of decidualization. PE-DSCs are significantly more responsive to sFlt1 inhibition by RA than are NT-DSC. Our finding of reduced RA levels in PE decidua, combined with studies by us and others that PE women show defective decidualization, may help explain elevated sFlt1 expression in maternal PE decidua.

Dysfunction of DNA damage-inducible transcript 4 in the decidua is relevant to the pathogenesis of preeclampsia.

Preeclampsia (PE) is a pregnancy-related disorder that occurs after 20 weeks of gestation and affects 3-5% of all human pregnancies worldwide. However, the pathogenesis of PE still remains poorly understood. A deficiency in decidualization is considered a contributing factor to the development of PE. The DNA damage inducible transcript 4 (DDIT4) gene encodes a protein whose main function is inhibiting mammalian target of rapamycin (mTOR) under stress, and several studies have demonstrated that its expression promotes tumor cell apoptosis. Our previous RNA-Seq results showed that DDIT4 is significantly decreased in the decidua of PE women. Here, we aimed to define the role of DDIT4 in human decidualization and its relationship with PE. The results indicated that DDIT4 was markedly decreased in the decidua of severe PE compared with those from uncomplicated pregnancies. The expression of DDIT4 in human endometrial stromal cell (hESC) line and primary hESCs was up-regulated during decidualization. Knockdown DDIT4 in hESCs and primary hESCs caused a significant reduction in the transcription of decidualization markers, insulin-like growth factor binding protein 1 (IGFBP1) and prolactin (PRL). In addition, silencing DDIT4 caused up-regulated p-mTOR and p-p70s6k and reduced apoptosis, whereas rapamycin, an inhibitor of mTOR, reversed the result of apoptosis. Moreover, the expression of cleaved-caspase 3 in severe PE was significantly lower than that of uncomplicated pregnancies, which was unfavorable for trophoblast invasion. Our data suggest that DDIT4 is critical for normal decidualization and the apoptosis of decidual cells. DDIT4 deficiency is likely involved in the development of PE.

Transcriptomic Profiling in Human Decidua of Severe Preeclampsia Detected by RNA Sequencing.

Maternal decidua plays a critical role in implantation and placentation. Impaired decidualization causes failed intravascular trophoblast invasion and inadequate placentation and then increases the risk of preeclampsia (PE). RNA sequencing (RNA-Seq) has achieved great advances in the characterization and quantification of transcriptomes; is a powerful tool for new transcript discovery, genome annotation, and expression profiling. In the present study, we conducted a RNA-Seq analysis to compare gene expression between decidua of PE (n = 3, early-onset severe PE, EOSPE; n = 3, late-onset severe PE, LOSPE) and normal pregnancies (n = 3). We revealed that decidual gene transcription profile was altered in severe PE and identified 293 key PE-related genes involved in 19 differentially regulated pathways relevant for the pathogenesis of PE, among which ENO2, PGK1, and HK2 involved in glycolysis/gluconeogenesis and HIF-1 signaling pathway which are all highly related with tumorigenesis and are significantly upregulated in cancer cells were severely inhibited in the decidua of PE. Moreover, we identified 22 core regulatory genes, including the newly identified pseudogenes BNIP3P1, HK2P1, and PGK1P1 that encode long non-coding RNA (lncRNA); interestingly, BNIP3/BNIP3P1, HK2/HK2P1, and PGK1/PGK1P1 appear in pairs in core genes. Subsequent analyses using quantitative PCR validated a portion of these results. This study may provide further insight into the mechanisms of PE and function as preventive, predictive, and therapeutic measures. Future functional studies are needed in order to accomplish a greater understanding of the mechanisms involved. J. Cell. Biochem. 119: 607-615, 2018. © 2017 Wiley Periodicals, Inc.

TGF-β3-induced miR-494 inhibits macrophage polarization via suppressing PGE2 secretion in mesenchymal stem cells.

Abnormal macrophage polarization at the maternal-fetal interface may contribute to the development of Preeclampsia (PE). The reason why macrophage polarization changed in PE is still unclear. Decidual mesenchymal stem cells (dMSCs) could regulate macrophage polarization. However, miRNA in dMSCs of PE were maladjusted. Therefore, we speculated that miRNA may affect dMSC-regulated macrophage polarization. In this study, we found that miR-494-overexpressed dMSCs inhibit M2 macrophage polarization and this inhibitory effect is mediated by miR-494-reduced PGE2 secretion. Furthermore, we proved that miR-494 is induced by TGF-β3. In summary, our findings suggest that the high expression of TGF-β3 in PE decidua stimulates miR-494 in dMSCs and attenuates the regulation of MSC switching the macrophage toward M2 type, contributing to an immune imbalance at maternal-fetal interface.

Anti-angiogenic collagen fragment arresten is increased from 16 weeks' gestation in pre-eclamptic plasma

IntroductionArresten and canstatin are endogenous anti-angiogenic factors derived from type IV collagen α-chains COL4A1 and COL4A2 respectively. While their functions are explored in cancer studies, little is known about their role in pregnancy. Pre-eclampsia (PE) is a common, serious hypertensive disorder of pregnancy that is characterised by systemic endothelial dysfunction. COL4A1 and COL4A2 are maternal PE susceptibility genes that have increased mRNA expression in PE decidua. Our study aim was to determine the levels of arresten and canstatin in plasma and decidua from PE and gestational age matched normotensive patients. MethodsPlasma was collected from normotensive (n = 44) and PE (n = 39) women during the second and third trimesters of pregnancy. Third trimester decidua was collected at delivery from normotensive and PE women (n = 4 each). Levels of arresten and canstatin were determined by Western immunoblotting. ResultsArresten levels were significantly increased in second and third trimester PE plasma, and in third trimester PE decidua (p < 0.05). Third trimester PE plasma arresten levels also significantly correlated with the need for MgSO4 treatment, where a 1.7 fold increase was observed in patients requiring MgSO4 treatment (p < 0.05). No significant change in canstatin levels was observed between normotensive and PE patients. DiscussionThis is the first study to report significant increases in the levels of collagen fragment arresten in PE plasma and decidua. Given its significant increase before the onset of clinical disease and associations with clinical severity in the third trimester, arresten may be a useful biomarker for predicting PE and monitoring its severity.

P34. Role of high-mobility group A1 protein in trophoblast invasion

Objectives Early in pregnancy, abundant neovascularization at the implantation site is important for normal placentation. For placental development and reduction of uterine vascular resistance, cytotrophoblasts must migrate to the decidua and the uterine myometrium, with subsequent invasion of the spiral arteries resulting in the spiral artery remodeling. Especially, extravillous trophoblasts (EVT) invade into the uterine decidual spiral arterioles and regulate the remodeling of these vessels for fetal blood supply. While, disturbed arterial remodeling can lead to the serious complications such as preeclampsia and fetal growth restriction. High-mobility group A1 protein (HMGA1) is known to play an important role in the proliferation of trophoblast; however, no specific function of HMGA1 has been reported for trophoblast invasion. The aim of this study was to evaluate the effect of HMGA1 on trophoblast invasion. Methods We investigated HMGA1 expressions in cytotrophoblast derived from preeclampsia model mouse, CD40-L mouse, using immunofluorescence analysis. Wound healing cell migration and matrigel invasion test was also performed using HTR-8/SVneo cells transfected with HMGA1 plasmid. Results HMGA1 was expressed in nucleus of trophoblasts derived from control mouse; in contrast, cytoplasmic expression was observed in CD40-L mouse. HMGA1 plasmid stimulated cell migration and invasion in HTR/SVneo cells; however, extranuclear translocation of HMGA1 suppressed cell migration and invasion. Conclusions Cellular localization disorder of HMGA1 resulted in a lowering of its invasions and migration. Transfer of HMGA1 in cytotrophoblast is crucial for the trophoblast invasion into decidua in preeclampsia.

81-OR

Objectives Preeclampsia (PE) is a common and serious pregnancy disorder with a strong genetic component. Numerous genetic studies performed, including our own, have yielded many susceptibility genes from distinct functional groups. Additionally, transcriptome profiling at the maternal-fetal interface has likewise yielded many differentially expressed genes. Often there is little overlap between the two approaches, although both are significantly informative with respect to PE. We have thus taken a novel approach by analysing pathways common to the susceptibility genes and the PE transcriptome. Methods Using Illumina Human Ht12v4 and Wg6v3 BeadChips, transcriptome profiling was conducted on n = 65 normotensive and n = 60 PE decidua basalis tissues collected at delivery. R software was used to pre-process transcript data for pathway analysis. Pathways were analysed and constructed using Ariadne Pathway Studio 9. We examined the following susceptibility genes: ACVR1, ACVR1C, ACVR2A, COL4A1, COL4A2, ERAP1, ERAP2, INHA, INHBB and LNPEP (Yong et al., 2014). Significance was set at p Results Major common targets/regulators of these susceptibility genes were AGT, IFNG, IL6, INHBA, SERPINE1, TGFB1 and VEGFA (Fig. 1). The top downstream pathway categories of susceptibility genes that were significantly altered in the PE decidual transcriptome were apoptosis and cell signalling. Conclusions Susceptibility genes from distinct functional groups have similar downstream pathways through common targets/regulators. Several of these pathways matched those altered in PE. This study contributes to a better understanding on how susceptibility genes may interact in the development of PE. With this knowledge, more targeted functional analyses of PE susceptibility genes in these key pathways can be performed to examine their contributions to the pathogenesis and severity of PE. Disclosures H.E. Yong: None. P.E. Melton: None. M.P. Johnson: None. K.A. Freed: None. B. Kalionis: None. P. Murthi: None. S.P. Brennecke: None. R.J. Keogh: None. E.K. Moses: None.

749: The impact of obesity on the disease course of preeclampsia

disease course of preeclampsia Emily Miller, Alan Peaceman Northwestern University, Maternal Fetal Medicine, Chicago, IL OBJECTIVE: Obesity is a well-established risk factor for the development of preeclampsia. Various possible causal etiologies have been proposed to explain this relationship. Insofar as these pathways toward preeclampsia may differ from the pathophysiology of preeclampsia in non-obese parturients, the clinical ramifications of the disease may differ. This study was designed to assess whether there is a difference in the clinical presentation of preeclampsia between obese and non-obese women. STUDY DESIGN: This is a retrospective cohort study of women who received a diagnosis of pre-eclampsia. Women were categorized as obese (pre-pregnancy BMI 30.0 kg/m2) or non-obese (pre-pregnancy BMI 30.0 kg/m2). Clinical data pertaining to their preeclampsia were abstracted from the chart, including gestational age at delivery, neonatal birth weight, blood pressure ranges, and lab values (creatinine, liver function tests, uric acid, platelet count, level of proteinuria). RESULTS: Of the 874 women included, 531 (61%) were obese. There were no statistically significant differences in maternal age, gestational age at delivery, BUN or creatinine levels, or amount of proteinuria between the two groups. While there was no difference in highest systolic blood pressure recorded, the non-obese cohort had a higher median maximum diastolic blood pressure [101 mm Hg (IQR 94107) v 98 mm Hg (IQR 92-104), p 0.001]. In addition, the non-obese group had a significantly increased risk of having an elevated AST (47% vs 32%, p 0.001), an elevated ALT (25% vs 18%, p 0.014), and thrombocytopenia (45% vs 30%). The non-obese women were twice as likely to develop both a platelet count of less than 100,000 and LFTs more than double of normal, consistent with the diagnosis of HELLP syndrome (6.7% v 2.8%, p 0.006). CONCLUSION: Of the women that were identified with preeclampsia, the majority were obese. However women with obesity and preeclampsia had lab abnormalities less frequently and also lower peak diastolic blood pressures. This suggests that the pathophysiology of preeclampsia may differ between obese and non-obese women. 750 Preeclampsia (PE) enhances NGAL expression at fetal-maternal interface: role of iron and hypoxia Errol Norwitz, Serkalem Tadesse, Zhonghua Tang, Seth Guller Tufts University School of Medicine, Obstetrics & Gynecology, Boston, MA, Tufts Medical Center, Obstetrics & Gynecology, Division of Maternal-Fetal Medicine, Boston, MA, Yale University School of Medicine, Obstetrics, Gynecology and Reproductive Sciences, New Haven, CT OBJECTIVE: PE is associated with both a substantial elevation in iron deposition and an aberrant maintenance of hypoxia at the maternalfetal interface. Indeed, increases in serum iron and iron transporting proteins such as neutrophil gelatinase-associated lipocalin (NGAL) have been proposed as potential antecedents of clinical symptoms in PE. This study investigates for the first time the possible role of iron, hypoxia, and NGAL in the etiology of PE using cultures of cytotrophoblasts (CTs) and decidual cells (DCs). STUDY DESIGN: To assess NGAL protein expression in PE, we analyzed placental tissues from normotensive (GA: 31 [30-32] wks) vs preeclamptic patients (GA: 31 [28-32] wks) by immunohistochemistry (IHC) and H-score analysis. CTs and DCs were isolated and purified from normal term placentas delivered by cesarean (n 8). To mimic the increased iron level and hypoxic environment in PE, cells were supplemented with iron citrate (25M) and exposed to normoxic (NMX; 20% O2) or hypoxic conditions (HPX; 2% O2 for 24h. NGAL protein and mRNA levels were measured by western blot and RTqPCR. RESULTS: (1) IHC revealed elevated NGAL protein expression in both extravillous trophoblast (EVT) and decidua in PE vs control (H-score: meanSEM): 375.175.7 vs 44.18.8 (p 0.003) and 286.352.7 vs 80.613.9 (p 0.001), respectively; (2) In CTs, iron supplementation increased NGAL expression under HPX (33.4-fold, p 0.0019), but not NMX; (3) In CTs, iron supplementation increased NGAL mRNA levels 3-fold under NMX but 175-fold under HPX (60-fold higher compared with HPX alone). Thus, HPX increased NGAL mRNA and protein expression in CTs in an iron-dependent manner. CONCLUSION: These data demonstrate that enhanced NGAL expression in EVT and decidua is a novel marker of PE. Further, our in vitro studies suggest that CT NGAL levels are elevated in PE through an iron-dependent, HPX-driven mechanism that is mediated at the level of gene transcription.

Decidual leucocyte populations in women with pre-eclampsia

Successful pregnancy is a carefully regulated balance of stimulatory and inhibitory immune mechanisms, dysregulation of which may generate harmful immune responses leading to pregnancy disorders. Evidence in the mouse suggests that the uterine vascular changes and altered trophoblast invasion in women with pre-eclampsia (PE) may be caused by altered maternal immune responses within the decidua. We tested the hypothesis that cytotoxic decidual leucocyte populations (CD3+, CD8+ and CD56+) are altered in women with PE. Placental bed biopsies from 12 women with PE [mean (SD) gestation 34.3 (4.5) weeks] and 12 women with normal pregnancies [36.8 (3.6) weeks] were studied using single immunohistochemical labelling (avidin–biotin complex method) and, where appropriate, double labelling (APAAP method) to detect CD3+, CD8+ and CD56+ leucocyte populations. Positively labelled decidual leucocytes were counted in five randomly selected fields. Compared with decidua from normal pregnancies, PE decidua contained fewer CD3+ cells [mean (SD) 40.3 (6.8) vs. 20.4 (1.9) cells per ×250 field, P < 0.01], CD8+ cells [34.8 (5.5) vs. 19.4 (2.6), P < 0.05] and CD56+ cells [29.0 (3.3) vs. 18.3 (1.9) cells per ×250 field, P < 0.01]. The reduction in each population of cytotoxic leucocytes was confirmed by double labelling.

Protein Carbonyls in Decidua and Placenta of Pre-eclamptic Women as Markers for Oxidative Stress

Enhanced levels of reactive oxygen species (ROS) and decreased levels of antioxidants may contribute to the development of pre-eclampsia. Protein carbonyls may be sensitive biomarkers for ROS-mediated protein damage, whereas the ferric reducing ability of plasma (FRAP) value may serve as marker for antioxidant capacity.Twenty-two healthy pregnant women and 50 patients with severe pre-eclampsia, 25 of whom with concurrent (haemolysis, elevated liver enzymes, low platelets) HELLP syndrome were investigated prospectively with respect to placental and decidual levels of protein carbonyls and FRAP antioxidant capacity. Measurements were evaluated using the Kruskal–Wallis or Mann–Whitney U -tests for comparison between groups, and the Wilcoxon signed-rank test for comparison within groups. Spearman's correlation tests were used for correlation analyses.Placental and decidual protein carbonyl levels were higher in pre-eclampsia with HELLP than in healthy pregnancy, whereas FRAP levels were lower. Differences between pre-eclampsia and healthy pregnancy showed the same trend, but did not reach statistical significance. In pre-eclampsia with HELLP syndrome carbonyl levels as well as FRAP levels were significantly higher in placenta as compared with decidua.Higher levels of protein carbonyls and lower antioxidant capacity in both placenta and decidua in pre-eclampsia with concurrent HELLP syndrome as compared to healthy pregnancy may indicate that considerable ROS-mediated damage occurs in this disorder.

Glutathione and Glutathione-related Enzymes in Decidua and Placenta of Controls and Women with Pre-eclampsia

Pre-eclampsia is a major complication of pregnancy with high morbidity and mortality rates. The aetiology is still unclear but impaired detoxification or enhanced levels of reactive (oxygen) metabolites may contribute to the development or maintenance of pre-eclampsia. Glutathione and glutathione-related enzymes, as one of the major detoxificating and free-radical scavenging systems, may play a role in controlling the disease.Seventeen normotensive pregnant women and 24 pre-eclamptic women were investigated prospectively with respect to placental and decidual levels of total glutathione (GSH), glutathione S-transferase activity (GST), selenium-dependent glutathione peroxidase (SeGPX) and total glutathione peroxidase activity (TGPX, both selenium- and non-selenium-dependent GPX). Decidual levels of glutathione and related enzymes were compared with placental levels, and the investigated parameters in pre-eclampsia were compared with those in normotensive pregnancy by the Mann–Whitney U -test. Clinical data were correlated with biochemical parameters by Spearman's correlation test.Glutathione levels were significantly higher in decidua as compared with placenta. Glutathione levels were elevated in pre-eclampsia and HELLP (haemolysis, elevated liver enzymes, low platelets) as compared to normotensive pregnancy for decidua and in the placenta of patients with pre-eclampsia only. Glutathione S-transferase activity was not different between the two groups. In the placenta of patients with pre-eclampsia+HELLP, total glutathione peroxidase activity was elevated versus controls. Selenium-dependent glutathione peroxidase activity was higher in decidua versus placenta and in decidua of pre-eclamptic versus control subjects.Enhanced glutathione concentrations and glutathione peroxidase activities were often found in placenta and decidua in pre-eclampsia, probably as a compensatory mechanism to prevent further damage by peroxides, (oxygen) radicals or other toxins in the placenta or in the feto–placental interface.

Pathogenetic implication of interleukin-2 expressed in preeclamptic decidual tissues: a possible mechanism of deranged vasculature of the placenta associated with preeclampsia.

The purpose of this study is to clarify whether IL-2 expressed in the decidua in preeclampsia affects the angiogenesis of the placenta. We investigated the angiogenic substances released from human trophoblasts obtained from early pregnancy that had been pretreated with either IL-2, non-activated lymphocytes from peripheral blood mononuclear cells (PBMCs), decidual lymphocytes, or these lymphocytes activated by lymphokine (LAK cells). Angiogenic activity was determined by evaluating the ability of growth-promotion of cultured human microvascular endothelial cells (HMvECs) using MTT assay. Trophoblasts pretreated with IL-2 or non-activated lymphocytes, irrespective of their origin, released angiogenic factor similar to those without pretreatment. However, trophoblasts pretreated with LAK cells released less angiogenic factor compared with those without pretreatment. Interleukin-2 (IL-2) expressed in preeclamptic decidua might reduce the angiogenic substances arising from trophoblasts by inducing LAK-cells from decidual lymphocytes, which might be relevant to deranged vasculature of the placenta, a characteristic histology in preeclampsia.

Quantitative Immunological Detection of Total Estrogen Receptor (Cytosolic and Nuclear) in Term Decidua of Preeclampsia: a Preliminary Study

Although progesterone and estrogens are essential for maintaining human pregnancy after implantation, few reports have investigated the localization of their specific receptors in different uterine cell types throughout pregnancy (1)(2). Wu et al. detected estrogen receptors (ER) in low concentrations in decidua in early pregnancy but not in term decidua (2). ER concentrations in term decidua of pregnancies with complications (in particular, preeclampsia) have not been investigated. Classical biochemical assay methods estimate ER either in cytosol fraction or in the high-salt extracted nuclei (3)(4)(5). These assays need two subcellular fractionation steps and two ER assays in two fractions: cytosol and nuclear extract. To measure total ER (cytosolic and nuclear) in one fraction using one single assay in decidual samples, we modified the assay method by using a homogenization buffer containing KCl. The effective KCl concentration for maximum ER extraction without inhibition of the ER enzyme immunoassay (EIA) ranged …

Decreased prolactin secretion by decidual tissue of pre-eclampsia in vitro.

In vitro prolactin (hPrl) secretion by explants of decidual tissue was studied in pre-eclampsia and normal controls. Our results indicate diminished hPrl production by decidua of pre-eclampsia as compared to the normal controls. Incubation of normal decidual tissue in the presence of serum obtained from pre-eclamptic patients did not induce an inhibitory effect on hPrl production.