transforming-growth-factor-beta and Pre-Eclampsia

Following embryo implantation, extravillous trophoblasts (EVTs) invade the maternal decidua to a certain extent during early pregnancy, which is critical for normal placentation and successful pregnancy in humans. Although sharing a similar protein structure, the transforming growth factor-β (TGF-β) superfamily members exert divergent functions in regulating EVT invasion, which contributes to a relative balance of TGF-β superfamily proteins in precisely modulating this process at the maternal-fetal interface during the first trimester of pregnancy. This review details recent advances in our understanding of the functions of TGF-β superfamily members and their corresponding receptors, signaling pathways, and downstream molecular targets in regulating human EVT invasion from studies using various in vitro or ex vivo experimental models. Also, the relevance of these discoveries about TGF-β superfamily members to adverse pregnancy outcomes is summarized. The application of 3D culture trophoblast organoids, single-cell sequencing, and microfluidic assays in EVT invasion studies will help better reveal the molecular mechanisms through which TGF-β superfamily members regulate human EVT invasion, shedding light on the development of innovative strategies for predicting, diagnosing, treating, and preventing adverse human pregnancy outcomes related to EVT invasion dysfunction.

Topics: Choriocarcinoma; Female; Humans; Pre-Eclampsia; Pregnancy; Receptors, Transforming Growth Factor beta; Transforming Growth Factor beta; Trophoblasts

Emerging data suggest that a trophoblast stem cell (TSC) population exists in the early human placenta. However, in vitro stem cell culture models are still in development and it remains under debate how well they reflect primary trophoblast (TB) cells. The absence of robust protocols to generate TSCs from humans has resulted in limited knowledge of the molecular mechanisms that regulate human placental development and TB lineage specification when compared to other human embryonic stem cells (hESCs). As placentation in mouse and human differ considerably, it is only with the development of human-based disease models using TSCs that we will be able to understand the various diseases caused by abnormal placentation in humans, such as preeclampsia. In this review, we summarize the knowledge on normal human placental development, the placental disease preeclampsia, and current stem cell model systems used to mimic TB differentiation. A special focus is given to the transforming growth factor-beta (TGFβ) family as it has been shown that the TGFβ family has an important role in human placental development and disease.

Topics: Cell Differentiation; Female; Humans; Placentation; Pre-Eclampsia; Pregnancy; Transforming Growth Factor beta; Trophoblasts

The etiology of preeclampsia - an abnormal placentation-mediated disease - is not fully understood; and there are very few biomarkers with which to predict and diagnose it. Early prediction and diagnosis of this pathology can lead to a significant improvement in maternal and perinatal outcomes. Since members of the transforming growth factor β superfamily influence placentation, and are released from the placenta into the maternal circulatory system, several studies have investigated the involvement of these cytokines in preeclampsia and the possibility of using their serum levels as biomarkers of the disease. In this review, we have summarized the reported relationships between the levels of this superfamily of cytokines and preeclampsia. The available information indicates that altered levels of some of these cytokines are involved in the pathogenesis and pathophysiology of preeclampsia, suggesting their likelihood of serving as predictive and diagnostic biomarkers of the disease.

Topics: Biomarkers; Female; Humans; Pre-Eclampsia; Pregnancy; Transforming Growth Factor beta

The role of angiogenic factors in the onset of clinical manifestations of preeclampsia was demonstrated in 2003 by the implication of sFlt-1, PlGF and VEGF, and in 2006 by the implication of soluble endoglin. Placental ischemia and inflammation observed in preeclampsia alter both the production and progression of angiogenic factors during pregnancy. During the first trimester, the combination of PlGF with clinical, biophysical and biological factors results in a better test than the conventional one. However, the clinical value of this method remains to be confirmed. During the second and third trimesters, the sFlt-1/PlGF ratio may be used, with or without pre-existing renal disease, for short-term prediction, diagnosis, and prognosis, and to evaluate the effectiveness of preeclampsia treatment. While a sFlt-1/PlGF ratio<38 and≤33, respectively, rules out the short-term onset and diagnosis of preeclampsia, a sFlt-1/PlGF ratio≥85 between 20 and 34 weeks of pregnancy and≥110 beyond 34 weeks of pregnancy confirms a diagnosis of preeclampsia. Angiogenic and non-angiogenic preeclampsia are identified by a sFlt-1PlGF≥85 and<85, respectively, with the risk of maternal and fetal complications at two weeks differing between the two. Similarly, a sFlt-1/PlGF ratio>665 and>205, respectively, is a good short-term predictor of adverse outcomes of early and late-onset preeclampsia. These values could be incorporated into future guidelines for better clinical management of preeclampsia.

Topics: Adult; Aspirin; Biomarkers; Endoglin; Endothelium, Vascular; Female; Humans; Immune Tolerance; Inflammation; Kidney Diseases; Kidney Transplantation; Membrane Proteins; Oxidative Stress; Placenta; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Pregnancy Outcome; Pregnancy Trimesters; Prognosis; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-1

Pre-eclampsia is a multi-system condition in pregnancy that is characterised by the onset of hypertension and proteinuria in women after the 20th week and it remains a leading cause of maternal and fetal mortality. Despite this the causative molecular basis of pre-eclampsia remains poorly understood. As a result, an intensive research effort has focused on understanding the molecular mechanisms involved in pre-eclampsia and using this information to identify new pre-symptomatic bio-markers of the condition. Activin A and its receptor, ACVR2A, have been extensively studied in this regard. Activin A is a member of the transforming growth factor (TGF)-β superfamily that has a wide range of biological functions depending on the cellular context. Recent work has shown that polymorphisms in ACVR2A may be a genetic risk factor for pre-eclampsia. Furthermore, both placenta and serum levels of Activin A are significantly increased in pre-eclampsia suggesting that Activin A may be a possible biomarker for the condition. Here we review the latest advances in this field and link these with new molecular data that suggest that the oxidative stress and pro-inflammatory cytokine production seen in pre-eclampsia may result in increased placental Activin A secretion in an attempt to maintain placental function.

Topics: Activins; Biomarkers; Clinical Trials as Topic; Cytokines; Female; Genetic Predisposition to Disease; Humans; Inflammation; Oxidative Stress; Placenta; Polymorphism, Genetic; Pre-Eclampsia; Pregnancy; Risk Factors; Signal Transduction; Transforming Growth Factor beta

T regulatory (Treg) cells are essential mediators of the maternal immune adaptation necessary for embryo implantation. In mice, insufficient Treg cell activity results in implantation failure, or constrains placental function and fetal growth. In women, Treg cell deficiency is linked with unexplained infertility, miscarriage, and pre-eclampsia. To devise strategies to improve Treg cell function, it is essential to define the origin of the Treg cells in gestational tissues, and the regulators that control their functional competence and recruitment. Male seminal fluid is a potent source of the Treg cell-inducing agents TGFβ and prostaglandin E, and coitus is one key factor involved in expanding the pool of inducible Treg cells that react with paternal alloantigens shared by conceptus tissues. In mice, coitus initiates a sequence of events whereby female dendritic cells cross-present seminal fluid antigens and activate T cells, which in turn circulate via the blood to be sequestered into the endometrium. Similar events may occur in the human genital tract, where seminal fluid induces immune cell changes that appear competent to prime Treg cells. Improved understanding of how seminal fluid influences Treg cells in women should ultimately assist in the development of new therapies for immune-mediated pathologies of pregnancy.

Topics: Abortion, Spontaneous; Animals; Dendritic Cells; Embryo Implantation; Endometrium; Female; Humans; Infertility; Isoantigens; Male; Mice; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Prostaglandins E; Semen; Seminal Plasma Proteins; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Endoglin (CD105) is a type III auxiliary receptor for the transforming growth factor beta (TGFβ) superfamily. Several lines of evidence suggest that endoglin plays a critical role in maintaining cardiovascular homeostasis. Seemingly disparate disease conditions, including hereditary hemorrhagic telangiectasia, pre-eclampsia, and cardiac fibrosis, have now been associated with endoglin. Given the central role of the TGFβ superfamily in multiple disease conditions, this review provides a detailed update on endoglin as an evolving therapeutic target in the management of cardiovascular disease.

Topics: Animals; Antigens, CD; Cardiovascular Diseases; Cardiovascular System; Endoglin; Female; Homeostasis; Humans; Ligands; Male; Pre-Eclampsia; Pregnancy; Receptors, Cell Surface; Signal Transduction; Telangiectasia, Hereditary Hemorrhagic; Transforming Growth Factor beta; Ventricular Remodeling

Conventional belief holds that an immune response to ejaculate antigens should interfere with fertilisation and establishment of pregnancy. However, emerging evidence now supports the opposing view-that insemination acts to activate maternal immune mechanisms exerting a positive effect on reproductive events. In a response well documented in rodents, semen triggers an influx of antigen-presenting cells into the female reproductive tract which process and present paternal ejaculate antigens to elicit activation of lymphocytes in the adaptive immune compartment. Transforming growth factor beta (TGFbeta), a cytokine present in abundance in seminal plasma, initiates this inflammatory response by stimulating the synthesis of pro-inflammatory cytokines and chemokines in uterine tissues. Lymphocyte activation is evident in lymph nodes draining the uterus and leads to hypo-responsiveness in T-cells reactive with paternal alloantigens. TGFbeta has potent immune-deviating effects and is likely to be the key agent in skewing the immune response against a Type-1 bias. Prior exposure to semen in the context of TGFbeta can be shown to be associated with enhanced fetal-placental development late in gestation. In this paper, we review the experimental basis for these claims and propose the hypothesis that, in women, the partner-specific protective effect of insemination in pre-eclampsia might be explained by induction of immunological hypo-responsiveness conferring tolerance to histocompatibility antigens present in the ejaculate and shared by the conceptus.

Topics: Animals; Antigen Presentation; Cytokines; Embryo Implantation; Female; Humans; Immune Tolerance; Inflammation Mediators; Male; Maternal-Fetal Exchange; Models, Immunological; Pre-Eclampsia; Pregnancy; Semen; Transforming Growth Factor beta; Trophoblasts

During early pregnancy, trophoblast differentiation occurs in an environment of relative low oxygen tension which is essential for normal embryonic and placental development. At around 10-12 weeks' gestation, when the intervillous space opens to maternal blood, there is an increase in Po(2). This increase correlates with the time of maximal trophoblast invasion into the maternal decidua, which allows extravillous trophoblast cells to access and remodel the maternal spiral arteries. Hypoxia Inducible Factor 1(HIF-1) is a transcription factor which activates gene transcription in response to varying oxygen concentration of cells. HIF-1 is a heterodimer composed of the inducible HIF-1alpha and the constitutively expressed HIF-1beta/ARNT subunits. Using villous explants, we have demonstrated that the oxygen-regulated events of early trophoblast differentiation are in part mediated by TGFbeta(3), an inhibitor of trophoblast differentiation, via HIF-1alpha. Pre-eclampsia is a disease of pregnancy that is characterized by shallow trophoblast invasion. Recently, we have reported that TGFbeta(3) is over-expressed in pre-eclamptic pregnancy and that its down-regulation restores invasive capability to trophoblast cells. Because TGFbeta(3) is downstream of HIF-1alpha, in the present study we investigated the expression of HIF-1alpha in pre-eclamptic placentae and age-matched controls using in situ hybridization and histochemical analyses. We found that HIF-1alpha mRNA and protein expression are abnormally elevated in pre-eclamptic placental tissue when compared to normal placental tissue. We conclude that pre-eclampsia may result from a developmental failure of oxygen to increase or of trophoblast cells to respond and/or sense an increase in oxygen. This will prevent the normal TGFbeta3 down-regulation and will lead to poor trophoblast invasion predisposing the pregnancy to pre-eclampsia.

Topics: Adult; Cell Differentiation; Chorionic Villi; DNA-Binding Proteins; Female; Gestational Age; Humans; Hypoxia-Inducible Factor 1; Hypoxia-Inducible Factor 1, alpha Subunit; Nuclear Proteins; Organ Culture Techniques; Oxygen; Pre-Eclampsia; Pregnancy; RNA, Messenger; Signal Transduction; Transcription Factors; Transforming Growth Factor beta; Transforming Growth Factor beta3; Trophoblasts

Pre-eclampsia and gestational hypertension are two common hypertensive disorders of pregnancy with pre-eclampsia accounting for high foetal and maternal morbidity and mortality rate. These disorders have an unknown aetiology and their hypertensive and end-organ pathophysiology may present too late in pregnancy. This makes the identification of early detection and differentiation markers vital. MicroRNAs have strongly been associated with pregnancy and their imbalance has been associated with the angiogenic dysregulation seen in pre-eclampsia. This study assesses the expression of pro- and antiangiogenic factors and their corresponding microRNAs in the maternal circulation of patients with pre-eclampsia and gestational hypertension.. We analyzed angiogenic factors expression (sEng, TGF-β, VEGF) normalized against housekeeping gene β-actin and microRNAs (miRs: 210, 29B, 126) normalized against miR U6, potentially associated with pre-eclampsia and gestational hypertension using the targeted qPCR technique. These analytes were examined from early-onset (<34 weeks) (EOPE) (n = 12), late-onset (>34 weeks) (LOPE) (n = 12) pre-eclampsia, gestational hypertension (GH) (n = 12) and two gestationally matched normotensive groups (NG1 and 2) (n = 12) each in South African women of African ancestry. Group comparisons of experimental vs. control groups were assessed using t-test analysis for significance and represented as fold change expression.. The relative expression in group comparisons showed significant (p < 0.05) fold change of VEGF, TGF-β, sEng and miR126 in the EOPE vs. NG1. The GH vs. NG1 exhibited significant changes in VEGF, TGF-β, miR126, miR210 and miR29B. The LOPE vs. NG2 showed significant relative expression in all the angiogenic factors (VEGF, TGF-β and sEng). The GH vs. NG2 showed significant expression in VEGF and miR29B. The LOPE vs. EOPE showed significant fold changes in VEGF and miR210. Finally, only the GH vs. EOPE showed significant differences in miR210 and miR29B (p < 0.05).. This study provides better insights into angiogenic factors and microRNAs specificity to the subtypes of gestational hypertensive disorders in pregnancy. Relative expression analysis of angiogenic factors and microRNAs showed possible novel characteristics of gestational hypertension, and potential common molecular and pathological profiles with pre-eclampsia. Furthermore, we postulate that sEng and miR29B could be early detection markers for pre-eclampsia and gestational hypertension, respectively.

Topics: Female; Humans; Hypertension, Pregnancy-Induced; MicroRNAs; Pre-Eclampsia; Pregnancy; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

We previously reported that cyclophilin A (CyPA) production is upregulated in preeclampsia (PE). Moreover, CyPA is known to induce PE-like features in pregnant mice and impair trophoblast invasiveness. In this study, we further illustrated the role of CyPA in PE. RNA-seq analysis, RT-qPCR, immunohistochemical (IHC) staining, and western blotting of mouse placentae revealed that CyPA increased the levels of extracellular matrix (ECM) proteins, such as collagen I and fibronectin, and activated the TGF-β/Smad3 signaling pathway. Additionally, CyPA inhibited the expression of genes involved in epithelial-mesenchymal transition (EMT) (e.g., E-cadherin, N-cadherin, and vimentin) in mouse placentae. We then constructed stable overexpressing and knock-down CyPA cell models (using HTR8/SVneo cells) to clarify the molecular mechanism. We found that CyPA regulated the levels of ECM-related proteins and the EMT process through the TGF-β/Smad3 pathway. We also identified SERPINH1 as a putative CyPA-binding protein, using liquid chromatography-electrospray mass spectrometry (LC-MS)/MS. SERPINH1 was found to be upregulated in the placentae of PE. Silencing SERPINH1 expression reversed the upregulation of ECM proteins and inhibition of the EMT process induced by the overexpression of CyPA. These findings revealed the functions of CyPA in the impaired invasiveness of trophoblasts in PE and indicated that CyPA and SERPINH1 may represent promising targets for the treatment of PE.

Topics: Animals; Cell Movement; Cyclophilin A; Epithelial-Mesenchymal Transition; Extracellular Matrix; Female; HSP47 Heat-Shock Proteins; Humans; Mice; Pre-Eclampsia; Pregnancy; Smad3 Protein; Transforming Growth Factor beta; Trophoblasts

Inadequate trophoblastic invasion is considered as one of hallmarks of preeclampsia (PE), which is characterized by newly onset of hypertension (>140/90 mmHg) and proteinuria (>300 mg in a 24-h urine) after 20 weeks of gestation. Accumulating evidence has indicated that long noncoding RNAs are aberrantly expressed in PE, whereas detailed mechanisms are unknown. In the present study, we showed that lncRNA Taurine upregulated 1 (TUG1) were downregulated in preeclamptic placenta and in HTR8/SVneo cells under hypoxic conditions, together with reduced enhancer of zeste homolog2 (EZH2) and embryonic ectoderm development (EED) expression, major components of polycomb repressive complex 2 (PRC2), as well as activation of Nodal/ALK7 signalling pathway. Mechanistically, we found that TUG1 bound to PRC2 (EZH2/EED) in HTR8/SVneo cells and weakened TUG1/PRC2 interplay was correlated with upregulation of Nodal expression via decreasing H3K27me3 mark at the promoter region of Nodal gene under hypoxic conditions. And activation of Nodal signalling prohibited trophoblast invasion via reducing MMP2 levels. Overexpression of TUG1 or EZH2 significantly attenuated hypoxia-induced reduction of trophoblastic invasiveness via negative modulating Nodal/ALK7 signalling and rescuing expression of its downstream target MMP2. These investigations might provide some evidence for novel mechanisms responsible for inadequate trophoblastic invasion and might shed some light on identifying future therapeutic targets for PE.

Topics: Activin Receptors, Type I; Cell Movement; Cell Proliferation; Female; Humans; Hypoxia; Matrix Metalloproteinase 2; Nodal Protein; Polycomb Repressive Complex 2; Pre-Eclampsia; Pregnancy; RNA, Long Noncoding; Taurine; Transforming Growth Factor beta; Trophoblasts

The Preeclampsia (PE) molecular mechanisms are not fully revealed and different biological processes are involved in the pathogenesis of PE. We aimed to evaluate adenosine and hypoxia-related signaling molecules in PE patients in the current study.. Decidua tissue and peripheral blood samples were taken from 25 healthy pregnant and 25 PE women at delivery time. CD39, CD73, and Hypoxia-inducible factor-alpha (HIF-α) were evaluated in mRNA and protein level using real-time PCR and western blotting techniques, respectively. Also, miR-30a, miR-206, and miR-18a expression were evaluated by real-time PCR. At last, secretion levels of IGF and TGF-β in the taken serum of blood samples were measured by ELISA.. Our results revealed that Expression of CD39 is decreased in PE cases versus healthy controls at mRNA and protein levels (p = 0.0003 for both). CD73 and HIF-α showed an increased level of expression in PE patients at RNA and protein status (p = 0.0157 and p < 0.0001 for protein evaluation of CD73 and HIF-α, respectively). The miRNA-30a (p = 0.0037) and miR-206 (p = 0.0113) showed elevated expression in the decidua of the PE group. The concentration of secreted IGF-1 (p = 0.0002) and TGF-β (p = 0.0101) in serum samples of PE cases compared to the healthy group were decreased.. In conclusion, our results showed that aberrant expression of molecules that are involved in ATP catabolism and the hypoxic conditions is observed in PE cases and involved in their hypertension and inflammation could be served as PE prognosis by more confirming in comprehensive future studies. miR-206 and miR-30a play a role by regulating CD39 and CD73 as molecules that are involved in ATP catabolism as well as regulating the production of IGF-1 in the process of hypertension, which is the main feature in patients with preeclampsia. On the other hand, decreased level of miR-18a lead to upregulation of HIF-1a, and the consequence condition of hypoxia increases hypertension and inflammation in these patients.

Topics: Adenosine Triphosphate; Decidua; Female; Humans; Hypertension; Hypoxia; Hypoxia-Inducible Factor 1, alpha Subunit; Inflammation; Insulin-Like Growth Factor I; MicroRNAs; Pre-Eclampsia; Pregnancy; Pregnant Women; RNA, Messenger; Transforming Growth Factor beta

Preeclampsia is one of the most serious pregnancy complications. It may be caused by immunological changes in the early placental microenvironment. The contents of small EVs may serve as biomarkers of pregnancy complications. Evidence suggests that the balance between T helper 17 (Th17) and regulatory T (Treg) cells are critical for preventing preeclampsia. The study recruited 39 pregnant women with preeclampsia and 127 healthy pregnant women. We assessed the levels of both Th17 and Treg cytokines (IL-10, IL-17, IL-21, IL-22, and TGF-β) in their plasma and small EVs. We found significant differences in the levels of all cytokines in the plasma between the two groups during the second trimester. We also observed significant differences between the two groups in the levels of EV-encapsulated cytokines IL-21, IL-22, and TGF-β, as well as in total small EVs, during the second trimester. The ROC analysis showed that the classification efficiency (AUC) of TGF-β in small EVs was 0.81. TGF-β had the best discriminant ability of all the single EV biomarkers tested, the cross-validation of the accuracy was 0.89. Th17 and Treg cytokines in plasma and small EVs may contribute to maternal immune activation and clarify the potential mechanisms of small EVs and cytokines in preeclampsia.

Topics: Biomarkers; Cytokines; Extracellular Vesicles; Female; Humans; Interleukin-10; Interleukin-17; Placenta; Pre-Eclampsia; Pregnancy; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta

Hofbauer cells (HBCs) are resident macrophages of the human placenta, regulating immune tolerance and tissue homeostasis. HBCs of a normal placenta (CTR) exhibit mainly an anti-inflammatory M2 phenotype. Under exaggerated chronic inflammation during pregnancy, as in preeclampsia (PE), a phenotypic switch towards M1 polarization has been proposed. PE, defined as maternally derived syndrome can be distinguished into two different entities: early-onset (EO) preeclampsia and late-onset (LO) preeclampsia. Although the clinical presenting characteristics overlap, both can be identified by biochemical markers, heritability, and different maternal and fetal outcomes. To date, no study has specifically investigated polarization and phenotype of EO- and LO-PE HBCs and looked at possible changes in HBC functionality. Primary HBCs were isolated from CTR and PE placentae. First,

Topics: Female; Humans; Inflammation; Macrophages; Placenta; Pre-Eclampsia; Pregnancy; Transforming Growth Factor beta

In preeclampsia (PE), human decidua mesenchymal stromal cells (hDMSCs) are exposed to abnormally high levels of oxidative stress and inflammatory factors circulating in the maternal blood. MicroRNAs (miRNAs) have been shown to have a significant impact on the differentiation, maturation and function of mesenchymal stromal cells (MSCs). Our aim in the present study is firstly to investigate differentially expressed miRNA levels to be used as a biomarker in the early detection of PE and secondly to investigate whether those differentially expressed miRNAs in hDMSCs have an effect on the pathogenesis of PE.. This study covers miRNA expression analysis of hDMSCs from 7 PE patient and 7 healthy pregnant women and is a preliminary study to investigate putative biomarkers. After cell culture and cell sorting, total RNA including miRNAs were isolated from hDMSCs. Let-7b-3p, let-7f-1-3p, miR-191-3p, miR-550a-5p, miR-33b-3p and miR-425-3p were used for miRNA analysis and U6 snRNA was used for normalization of the samples. MiRNA analysis was performed by droplet digital polymerase chain reaction (ddPCR) method and obtained results were evaluated statistically.. As a result of the analysis, it was observed that the levels of hsa-miR-33b-3p significantly (AUC: 0.93, p = 0.04, fold change: 4.5) increased in hDMSC of PE patients compared to healthy controls. However, let-7b-3p, let-7f-1-3p, miR-191-3p, miR-550a-5p, and miR-425-3p were not considered as significant because they did not meet the p < 0,05 requirement.. Within the scope of the study, it is predicted that miR-33b-3p (p = 0.004, AUC = 0.93) can be used as a biomarker in detecting PE.

Topics: Adult; Cesarean Section; Decidua; Female; Humans; Mesenchymal Stem Cells; MicroRNAs; Polymerase Chain Reaction; Pre-Eclampsia; Pregnancy; Transforming Growth Factor beta

Preeclampsia (PE) is a leading cause of stroke and cognitive impairment in the offspring. Melatonin is involved in the outcome of normal pregnancy. Its receptors are widespread in the embryo. This study aimed to investigate the fetal neuroprotective effect of melatonin in experimentally induced PE. After induction of pregnancy in 18 female rats, they were divided into three equal groups. PE was induced in groups II and III by injection of deoxycorticosterone acetate and drinking isotonic saline. Melatonin was supplied to group III orally (10 mg/kg body weight) throughout pregnancy. Pregnancy was terminated on day 20, and macroanatomical investigation of three fetuses from each pregnant rat and their placentae was performed. Placental and brain homogenates were analyzed for malondialdehyde (MDA), placental growth factor (PLGF), tumor necrosis factor-α (TNF-α), and brain transforming growth factor-β (TGF-β). Histopathological analysis of fetal brain sections was performed. Melatonin improved placental, fetal, and brain weight; significantly reduced fetal death rate; significantly increased PLGF, placental and brain superoxide dismutase, and brain TGF-β; and significantly decreased placental TNF-α and brain MDA. Brain micromorphological study found normal glial cells and neuropil in the melatonin-treated group and a loss of neuronal cell outlines with an accumulation of cellular debris in the untreated group. In conclusion, melatonin approximately showed a neuroprotective activity by managing PE-induced oxidative stress in the placenta and fetal cerebral cortex of rats.

Topics: Animals; Brain; Disease Models, Animal; Female; Malondialdehyde; Melatonin; Oxidative Stress; Placenta Growth Factor; Pre-Eclampsia; Pregnancy; Rats; Superoxide Dismutase; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Preeclampsia (PE) is commonly considered as a placental disorder in pregnancy. Until now, the etiology and pathological mechanism of PE have remained ambiguous. Although PE can lead to a variety of maternal and infant complications, there are still no effective treatments. This study aimed to explore the correlation between the novel polypeptide COL-4A1 and PE, and to identify the underlying mechanism by which this polypeptide may function and to explore new therapeutic targets for PE. A rat model of PE was established and used to verify the function of the polypeptide COL-4A1 in vivo. Additionally, human umbilical vascular endothelial cells (HUVECs) were cultured with or without COL-4A1 and TNF-α (20 ng/ml). Cell Counting Kit-8 (CCK-8), wound-healing, Transwell and tube formation assays were used to evaluate cell proliferation, migration and angiopoiesis. RNA sequencing and mass spectrometry were conducted to explore the underlying downstream mechanism of COL-4A1. In vivo, COL-4A1 increased blood pressure and elevated the risk of fetal growth restriction (FGR) which was induced by lipopolysaccharide (LPS) in the rat model. In vitro, COL-4A1 significantly inhibited the proliferation and migration of HUVECs. After culture with COL-4A1, compared to control group the adhesive ability and level of reactive oxygen species (ROS) were enhanced and tube formation ability was decreased. Furthermore, Western blotting (WB) and pull-down assays were conducted to explore the underlying mechanism by which COL-4A1 functions, and the TGF-β/PI3K/AKT pathway was identified as the potential pathway involved in its effects. In summary, these results revealed that the polypeptide COL-4A1 caused PE-like symptoms in cells and a rat model. Through the TGF-β/PI3K/AKT pathway, COL-4A1 interferes with the pathogenesis of PE. Thus COL-4A1 is expected to become a potential target of PE, providing a basis for exploring the treatment of PE.

Topics: Animals; Cell Movement; Cell Proliferation; Collagen Type IV; Female; Fetal Growth Retardation; Human Umbilical Vein Endothelial Cells; Humans; Hypertension; Male; Peptides; Phosphatidylinositol 3-Kinases; Pre-Eclampsia; Pregnancy; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Signal Transduction; Transforming Growth Factor beta

Endoglin is a membrane glycoprotein primarily expressed by the vascular endothelium and involved in cardiovascular diseases. Upon the proteolytic processing of the membrane-bound protein, a circulating form of endoglin (soluble endoglin, sEng) can be released, and high levels of sEng have been observed in several endothelial-related pathological conditions, where it appears to contribute to endothelial dysfunction. Preeclampsia is a multisystem disorder of high prevalence in pregnant women characterized by the onset of high blood pressure and associated with increased levels of sEng. Although a pathogenic role for sEng involving hypertension has been reported in several animal models of preeclampsia, the exact molecular mechanisms implicated remain to be identified. To search for sEng-induced mediators of hypertension, we analyzed the protein secretome of human endothelial cells in the presence of sEng. We found that sEng induces the expression of BMP4 in endothelial cells, as evidenced by their proteomic signature, gene transcript levels, and BMP4 promoter activity. A mouse model of preeclampsia with high sEng plasma levels (

Topics: Animals; Bone Morphogenetic Protein 4; Carrier Proteins; Endoglin; Endothelial Cells; Female; Humans; Hypertension; Mice; Mice, Inbred C57BL; Mice, Transgenic; Pre-Eclampsia; Pregnancy; Proteomics; Transforming Growth Factor beta; Up-Regulation

Background Preeclampsia (PE) is a multisystem disease and is still among the leading causes of maternal and neonatal morbidity and mortality. Inadequate trophoblast invasion plays a key role in the PE pathogenesis. The proliferation, migration, and invasion of extravillous trophoblasts (EVTs) is primarily controlled by the decidua-derived transforming growth factor beta (TGF-β) and decorin. In this study, we aimed to investigate the clinical utility of serum decorin levels measured in the 11th to 14th gestational weeks to predict preeclampsia during the following weeks of gestation. Materials and Methods A total of 600 pregnant women, whose age and gestational age ranged from 18 to 40 years and 11 to 14 weeks, were included. Venous blood samples were obtained and stored at -80 °C. Subsequently, the patients who developed preeclampsia and healthy controls with a similar body mass index were identified and their first-trimester blood samples were analyzed for serum decorin levels. Results The mean serum decorin level was 8.76 ± 6.88 ng/mL for the PE group while 9.75 ± 9.82 ng/mL for the control group. No statistically significant difference was found between the two groups (p=0.838). Conclusion We observed that the serum decorin levels during the 11th to 14th weeks of gestation showed no predictive value for preeclampsia in pregnant women. However, more accurate conclusions about the clinical utility of decorin as a biomarker of preeclampsia would require further studies with larger samples including more patients with EOS-PE.

Topics: Adult; Correlation of Data; Decidua; Decorin; Female; Gestational Age; Humans; Placenta; Pre-Eclampsia; Predictive Value of Tests; Pregnancy; Pregnancy Trimester, First; Prognosis; Risk Assessment; Transforming Growth Factor beta; Trophoblasts

Preeclampsia, especially early-onset severe preeclampsia is one of the leading causes of maternal and fetal morbidity and mortality. Although it has been well known that the pathophysiology of early-onset severe preeclampsia begins with abnormal placentation and aberrant activation of TGF-β signaling inhibits trophoblast cell invasion, the mechanisms underlying dysregulation of TGF-β signaling in early-onset severe preeclampsia remain elusive to date. Here, we revealed that induction of TGFBR1/TGF-β signaling mediated by DNMT3A downregulation plays a critical role in early-onset severe preeclampsia. Our results show that DNMT3A downregulation elevates TGFBR1 expression in trophoblast cells. Moreover, inhibition of TGFBR1 and TGF-β/Smad signaling can rescue the deficiencies of trophoblast cell migration and invasion caused by DNMT3A knockdown. Mechanistically, DNMT3A suppresses the transcription of TGFBR1 through recruiting EZH2 to its promoter but not changing DNA methylation of TGFBR1 promoter. In human samples, we detected lowly expressed DNMT3A, highly expressed TGFBR1 and hyperactivation of TGF-β/Smad signaling in decidua-embedded extravillous trophoblasts in early-onset severe preeclampsia, which provides the clinical evidence for the correlation between DNMT3A and TGFBR1. Collectively, our findings demonstrate that DNA methylation-independent induction of TGFBR1 mediated by DNMT3A downregulation is relevant to the development of early-onset severe preeclampsia.

Topics: Adult; Cell Line; DNA (Cytosine-5-)-Methyltransferases; DNA Methylation; DNA Methyltransferase 3A; Down-Regulation; Enhancer of Zeste Homolog 2 Protein; Female; Humans; Pre-Eclampsia; Pregnancy; Promoter Regions, Genetic; Receptor, Transforming Growth Factor-beta Type I; Signal Transduction; Smad Proteins; Transforming Growth Factor beta; Trophoblasts

To explore the protective effect of micro ribonucleic acid (miR)-210 on cranial nerves in rats with preeclampsia (PE) by regulating the transforming growth factor-β (TGF-β) signaling pathway.. A total of 36 pregnant Sprague-Dawley rats were randomly divided into normal group (n=12), model group (n=12), and miR-210 mimics group (n=12). The rats were fed normally in the normal group. In the latter two groups, the PE model was established, followed by injection of normal saline or miR-210 mimics via the caudal vein, respectively. The above intervention lasted until 20 d of gestational age in pregnant rats. Then, the systolic blood pressure of the caudal vein was measured. The relative levels of Caspase3, phosphorylated TGF-β (p-TGF-β), and miR-210 were detected via immunohistochemistry, Western blotting, and quantitative Polymerase Chain Reaction (qPCR). Cell apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay.. The systolic blood pressure of the caudal vein significantly increased in the other two groups compared with that in the normal group (p<0.05), while it significantly decreased in the miR-210 mimics group compared with that in the model group (p<0.05). The results of immunohistochemistry showed that the positive expression of Caspase3 significantly rose in the other two groups compared with that in the normal group (p<0.05), while it remarkably declined in miR-210 mimics group compared with that in the model group (p<0.05). The results of Western blotting revealed that the protein expression of p-TGF-β was evidently higher in the other two groups than that in the normal group (p<0.05), while it was evidently lower in the miR-210 mimics group than that in the model group (p<0.05). Moreover, it was found via qPCR that the other two groups had remarkably lower relative expression of miR-210 than normal group (p<0.05), while miR-210 mimics group had remarkably higher relative expression of miR-210 than the model group (p<0.05). According to the results of TUNEL assay, the apoptosis rate markedly increased in the other two groups compared with that in the normal group (p<0.05), while it markedly decreased in the miR-210 mimics group compared with that in the model group (p<0.05).. MiR-210 inhibits apoptosis via suppressing the TGF-β signaling pathway, thereby exerting a protective effect on cranial nerves in PE rats.

Topics: Animals; Apoptosis; Cranial Nerves; Female; MicroRNAs; Pre-Eclampsia; Pregnancy; Protective Agents; Rats; Rats, Sprague-Dawley; Signal Transduction; Transforming Growth Factor beta

An association between preeclampsia and (pro)renin was recently reported. Intracellular signaling of the (pro) renin receptor [(P)RR] increases the expressions of TGF-β and PAI-1. In this study we sought to clarify the involvement of (pro)renin in the pathogenesis of preeclampsia via the intracellular signaling of (P)RR on preeclampsia placentas. Activated (pro)renin plasma concentrations were compared between pregnant women with (n=15) and without (n=28) preeclampsia. The placentas were immunohistochemically evaluated with anti-HIF-1α and anti-(P)RR antibodies. HTR-8/SVneo cells were cultured under hypoxic conditions and treated with human recombinant (pro)renin. The mRNA expressions of HIF-1α, (P)RR, PAI-1, TGF-β, and ET-1 were also examined by real-time RCR. The activated (pro)renin plasma concentration was significantly higher in the third vs. the second trimester in the preeclampsia patients. HIF-1α and (P)RR expressions were significantly increased in the preeclampsia placentas. The mRNA expressions of PAI-1, TGF-β, and ET-1 were significantly increased in the experiments using recombinant (pro)renin vs. hypoxic conditions. (P)RR expression in preeclampsia placentas is increased by persistent hypoxia through the second and third trimesters, and PAI-1, TGF-β, and ET-1 production is increased via (P)RR. Our results suggest that ET-1 production via the intracellular signaling of (P)RR is important in the pathogenesis of preeclampsia.

Topics: Adult; Cells, Cultured; Endothelin-1; Female; Humans; Plasminogen Activator Inhibitor 1; Pre-Eclampsia; Pregnancy; Prorenin Receptor; Receptors, Cell Surface; Signal Transduction; Transforming Growth Factor beta

Decidual natural killer (dNK) cells are the predominant lymphocytes accumulated at the maternal-fetal interface. Regulatory mechanism of dNK cells in preeclampsia, a gestational complication characterized by high blood pressure and increased proteinuria occurring after 20 weeks pregnancy, is not completely understood.. Multi-parameter flow cytometry is applied to investigate the phenotype and function of dNK cells freshly isolated from decidual samples or conditionally cultured by TGFb stimulation.. Our findings suggest that elevated decidual TGFb1 supresses the activation of specific subsets of dNK which in turn contributes to the uteroplacental pathology associated with the onset of preeclampsia.

Topics: Cells, Cultured; Culture Media, Conditioned; Decidua; Down-Regulation; Female; Flow Cytometry; Humans; Interferon-gamma; Interleukin-8; Killer Cells, Natural; Lysosomal-Associated Membrane Protein 1; Pre-Eclampsia; Pregnancy; Signal Transduction; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Transforming Growth Factor beta1

Preeclampsia is a pregnancy-specific disorder that is a major cause of maternal and fetal morbidity and mortality with a prevalence of 6-8% of pregnancies. Although impaired trophoblast invasion in early pregnancy is known to be closely associated with preeclampsia, the underlying mechanisms remain elusive. Here we revealed that lysyl oxidase (LOX) and LOX-like protein 2 (LOXL2) play a critical role in preeclampsia. Our results demonstrated that LOX and LOXL2 expression decreased in preeclamptic placentas. Moreover, knockdown of LOX or LOXL2 suppressed trophoblast cell migration and invasion. Mechanistically, collagen production was induced in LOX- or LOXL2-downregulated trophoblast cells through activation of the TGF-β1/Smad3 pathway. Notably, inhibition of the TGF-β1/Smad3 pathway could rescue the defects caused by LOX or LOXL2 knockdown, thereby underlining the significance of the TGF-β1/Smad3 pathway downstream of LOX and LOXL2 in trophoblast cells. Additionally, induced collagen production and activated TGF-β1/Smad3 were observed in clinical samples from preeclamptic placentas. Collectively, our study suggests that the downregulation of LOX and LOXL2 leading to reduced trophoblast cell migration and invasion through activation of the TGF-β1/Smad3/collagen pathway is relevant to preeclampsia. Thus, we proposed that LOX, LOXL2, and the TGF-β1/Smad3/collagen pathway can serve as potential markers and targets for clinical diagnosis and therapy for preeclampsia.

Topics: Amino Acid Oxidoreductases; Biomarkers; Cell Line; Cell Movement; Collagen; Female; Gene Knockdown Techniques; Humans; Pre-Eclampsia; Pregnancy; Protein-Lysine 6-Oxidase; RNA, Messenger; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta; Trophoblasts

To measure the peripheral blood mononuclear cells (PBMCs) mRNA expression of placental growth factor (PlGF), Transforming growth factor beta (TGF-β), and soluble Endoglin (sEng) in the blood of preeclamptic and normotensive pregnant women.. Cross-sectional analytical study.. Department of Physiology and Cell Biology, University of Health Sciences, Lahore, from November 2016 to April 2018.. The study included 50 normotensive and 57 preeclamptic patients (18-40 years of age), all in the third trimester of pregnancy. The preeclamptic group was further divided into early-onset preeclampsia (EOP) and late-onset preeclampsia (LOP). Blood samples from patients and healthy controls were collected and mRNA expression was measured (18 patients and 18 controls) by real time PCR. Statistical analyses were done using SPSS (version 22). The values were considered significant at 0.05 level of significance.. The PBMCs mRNA expression of PlGF, TGF- and sEng were significantly different between the preeclampsia and control group (p<0.001). A significant decrease in expression of TGF- was observed in LOP group compared to controls (p<0.001); whereas, the difference in the expression of EOP compared to controls was not significant (p=0.12). Similar to TGF-, the expression of PlGF was significantly decreased among EOP and LOP compared to controls. Detailed analysis of sEng showed significantly increased expression in both EOP and LOP as compared to healthy group (p<0.001).. There is a significant difference in extra-placental expression of PlGF, and sEng in preeclampsia.

Topics: Adolescent; Adult; Biomarkers; Cells, Cultured; Cross-Sectional Studies; Endoglin; Female; Follow-Up Studies; Humans; Leukocytes, Mononuclear; Membrane Proteins; Pakistan; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Third; Real-Time Polymerase Chain Reaction; Reference Values; Risk Assessment; RNA, Messenger; Transforming Growth Factor beta; Young Adult

Pre-eclampsia is a common pregnancy complication, affecting 5-8% of pregnancies worldwide. The specific mechanism of pre-eclampsia remains unclear.. In this study, we aimed to apply bioinformatics approach to reveal related pathways or genes involving in the development of pre-eclampsia.. The gene expression profiles of GSE9984 and GSE4707 were downloaded from the Gene Expression Omnibus database. Differentially expressed genes analysis was performed by GEO2R. The Database for Annotation, Visualization and Integrated Discovery (DAVID) was applied to analyze the functional enrichment, gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway of the differentially expressed genes. Gene Set Enrichment Analysis (GSEA) was conducted using the software GSEA v3.0. Protein-protein interaction (PPI) relationships were evaluated by the Search Tool for the Retrieval of Interacting Genes (STRING) and network visualization was constructed by the Cytoscape. Cell count kits-8 (CCK-8), transwell migration assay and tube formation assay were performed.. A total of 160 common differentially expressed genes were extracted. Transforming growth factor (TGF) beta signaling pathway was shown to be notable in the development of pre-eclampsia. ENG, a key gene of TGF-β signaling pathway, inhibited the proliferation, migration and invasion of both HTR-8/SVneo cells and human umbilical vein endothelial cells (HUVECs), and additionally suppressed the capillary formation of HUVECs.. Bioinformatics approach combined with cell experiments in this study revealed that TGF-β signaling pathway was critical for the development of pre-eclampsia, and efficient biomarkers underlying this pathway need to be further investigated.

Topics: Cell Line; Cell Movement; Cell Proliferation; Computational Biology; Databases, Genetic; Female; Human Umbilical Vein Endothelial Cells; Humans; Pre-Eclampsia; Pregnancy; Protein Interaction Maps; Signal Transduction; Transcriptome; Transforming Growth Factor beta

A growing body of evidence suggests that the dysregulation of long noncoding RNA is increasingly linked to many human diseases. Maternally expressed gene 3 ( MEG3) is one such gene thought to be affected. In the placenta of patients with preeclampsia, there is reduced expression of MEG3; however, its role and the mechanism involved are not clear. Therefore, we examined the expression of MEG3, epithelial-mesenchymal transition (EMT) markers (E-cadherin and N-cadherin), and TGF-β/smad signaling pathway genes ( TGF-β1, smad3, and smad7) in the placental tissues of 20 patients with preeclampsia and 20 healthy patients. We further observed the impact of MEG3 on the invasion and migration functions of human trophoblast cells and the effects on EMT and TGF-β/smad signaling pathways in an Human trophoblast cell-8 (HTR-8)Vneo cell line. The expression of MEG3 was lower in tissues from patients with preeclampsia having an EMT decline, as well as a messenger RNA expression of smad7. The expression of TGF-β1 and smad3 were higher in patients with preeclampsia. In HTR-8/SVneo cells with overexpressed MEG3, the invasion and migration functions were enhanced and accompanied by higher EMT and a significantly increased expression of smad7. Our data indicate that MEG3 is closely associated with the pathogenesis of preeclampsia and thus associated with changes in the EMT of placental trophoblast cells. These results indicate that MEG3 regulation of trophoblast cell EMT via the TGF-β pathway inhibitor smad7 may be the molecular mechanism involved in the pathogenesis of preeclampsia.

Topics: Adult; Cadherins; Cell Movement; Epithelial-Mesenchymal Transition; Female; Humans; Pre-Eclampsia; Pregnancy; RNA, Long Noncoding; Signal Transduction; Smad Proteins; Transforming Growth Factor beta; Trophoblasts

Preeclampsia (PE) can be classified into early-onset (<34 weeks of gestation) and late-onset (>34 weeks of gestation) subtypes. Soluble endoglin, an auxiliary receptor for transforming growth factor (TGF)-β ligands, is increased in PE circulation and believed to inhibit TGF-β action by sequestering the ligands. However, soluble endoglin, with a low affinity to TGF-β ligands, has been demonstrated to have little effect by itself on TGF-β action.. We examined whether multiple soluble TGF-β receptors are elevated in PE circulation and whether they synergistically block TGF-β signaling.. TGF-β receptors were measured using enzyme-linked immunosorbent assay in sera collected from preeclamptic pregnancies and gestation-age-matched controls. TGF-β signaling was assessed using an in vitro bioassay and a tube formation assay.. TGF-β type I, II, and III receptors were all identified in pregnant serum; all were substantially elevated in early-onset but not late-onset PE. Endoglin was increased in both subtypes. At the greatest concentrations detected in PE, none of these soluble TGF-β receptors alone, including endoglin, inhibited TGF-β signaling. However, when all four soluble receptors were present, signaling of both TGF-β1 and TGF-β2 was substantially reduced. Removal of any one of these soluble receptors alleviated TGF-β1 inhibition; however, removal of soluble TGFβRIII was necessary to relieve TGF-β2 inhibition.. Multiple soluble TGF-β receptors are present in pregnant circulation and elevated in early-onset PE; they synergistically inhibit TGF-β signaling, which might be more likely to occur in early-onset than late-onset PE. Reducing soluble TGFβRIII, rather than endoglin, would be more effective in alleviating the inhibition of both TGF-β1 and TGF-β2 signaling in PE.

Topics: Adult; Case-Control Studies; Endoglin; Enzyme-Linked Immunosorbent Assay; Female; Gestational Age; HEK293 Cells; Human Umbilical Vein Endothelial Cells; Humans; In Vitro Techniques; Pre-Eclampsia; Pregnancy; Protein Serine-Threonine Kinases; Proteoglycans; Receptor, Transforming Growth Factor-beta Type I; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Signal Transduction; Transforming Growth Factor beta; Young Adult

Preeclampsia (PE), a condition during pregnancy that involves high blood pressure and proteinuria, is potentially fatal to both mother and child. PE currently has no known etiology or cure but has been tied to poor placental trophoblast cell migration. Increased levels of the toxic metal cadmium (Cd) have been associated with increased risk of developing PE, as well as miRNA-associated regulation of the transforming growth factorbeta (TGF-β) pathway. Signal reprogramming of the TGF-β pathway via epigenetic mechanisms is hypothesized to modify placental trophoblast function. In the present study we investigated the role of increased and decreased signaling of the TGF-β pathway in relation to Cd-induced reduction in cellular migration in JEG3 trophoblast cells. Furthermore, the role of a miR-26a as a molecular mediator of placental trophoblast migration was confirmed. The results demonstrate that increased expression of miR-26a and decreased signaling of the TGF-β pathway increase placental cell migration. These findings have relevance for mechanistic understanding of the underpinnings of poor placentation associated with PE.

Topics: Cadmium; Cell Line; Cell Movement; Female; Humans; MicroRNAs; Placenta; Pre-Eclampsia; Pregnancy; Signal Transduction; Transforming Growth Factor beta; Trophoblasts

Pre-eclampsia (PE), a hypertensive disorder of pregnancy, is detrimental to both mother and foetus. There is currently no effective treatment, but we have shown that Sildenafil Citrate (SC) improve various foetal outcomes in N

Topics: Animals; Biomarkers; Blood Pressure; Disease Models, Animal; Female; Gene Expression Regulation; Inflammation; Interferon-gamma; Neovascularization, Physiologic; NG-Nitroarginine Methyl Ester; Nitric Oxide; Pre-Eclampsia; Pregnancy; Rats; Rats, Sprague-Dawley; RNA, Messenger; Sildenafil Citrate; Time Factors; Transforming Growth Factor beta; Uterus; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-1

To assess circulating levels of transforming growth factor (TGF)-β superfamily members and their soluble receptors in hypertensive disorders of pregnancy, and to investigate associations with clinical manifestations.. A retrospective study was conducted using data for women admitted to a center in China for delivery between May 2011 and April 2013. Women with severe pre-eclampsia, mild pre-eclampsia, and gestational hypertension were included, along with a control group. Serum levels of activin A, inhibin A, TGF-β1, soluble endoglin (sEng), and soluble betaglycan (sBG) were measured.. Women with severe pre-eclampsia (n = 17) had higher mean levels of activin A (23.5±2.1 μg/L), inhibin A (1.7±0.2 μg/L), sEng (32.1±3.2 μg/L), and sBG (84.1±9.4 μg/L) than did normotensive controls (n = 18), women with gestational hypertension (n = 15), and those with mild pre-eclampsia (n = 14; all P<0.05). Women with early-onset pre-eclampsia (n = 13) had higher levels of these serum markers than did preterm normotensive controls (n = 8; all P<0.001). Women with severe or early-onset pre-eclampsia had the lowest TGF-β1 levels. Activin A, inhibin A, sEng, and sBG levels were positively correlated with mean arterial pressure and proteinuria (all P<0.01).. Pre-eclampsia is associated with an imbalance of members of the TGF-β superfamily and their soluble receptors, which might contribute to the development of pre-eclampsia and help to predict onset and severity.

Topics: Activins; Adult; Endoglin; Female; Humans; Hypertension, Pregnancy-Induced; Inhibins; Pre-Eclampsia; Pregnancy; Proteoglycans; Receptors, Transforming Growth Factor beta; Retrospective Studies; Transforming Growth Factor beta; Transforming Growth Factor beta1

TGFβ has been implicated in preeclampsia, but its intracellular signaling via phosphorylated mothers against decapentaplegic (SMADs) and SMAD-independent proteins in the placenta remains elusive. Here we show that TGFβ receptor-regulated SMAD2 was activated (Ser(465/467) phosphorylation) in syncytiotrophoblast and proliferating extravillous trophoblast cells of first-trimester placenta, whereas inhibitory SMAD7 located primarily to cytotrophoblast cells. SMAD2 phosphorylation decreased with advancing gestation, whereas SMAD7 expression increased and shifted to syncytiotrophoblasts toward term. Additionally, we found that the TGFβ SMAD-independent signaling via partitioning defective protein 6 (PARD6)/Smad ubiquitylation regulatory factor was activated at approximately 10-12 weeks of gestation in cytotrophoblast and extravillous trophoblast cells comprising the anchoring column. Placentae from early-onset, but not late-onset, preeclampsia exhibited elevated SMAD2 phosphorylation and SMAD7 levels. Whereas PARD6 expression increased and SMURF1 levels decreased in preeclamptic placentae, their association increased. SMAD2 phosphorylation by TGFβ in villous explants and BeWo cells resulted in a reduction of Glial cell missing-1 (GCM1) and fusogenic protein syncytin-1 while increasing cell cycle regulators cyclin E-1 (CCNE1) and cyclin-dependent kinase 4. SMAD7 abrogated the proliferative effects of TGFβ. CCNE1 levels were increased in preeclamptic placentae, whereas GCM1 was markedly reduced. In addition, TGFβ treatment increased the association of PARD6 and SMURF1 and down-regulated Ras homolog gene family, member A (RHOA) GTPase in JEG3 cells. In a wound assay, TGFβ treatment increased the association of PARD6 and SMURF1 and triggered JEG3 cell migration through increased cellular protrusions. Taken together, our data indicate that TGFβ signaling via both SMAD2/7 and PARD6/SMURF1 pathways plays a role in trophoblast growth and differentiation. Altered SMAD regulation of GCM1 and CCNE1 and aberrant expression/activation of PARD6/SMURF1 may contribute to the pathogenesis of preeclampsia by affecting cellular pathways associated with this disorder.

Topics: Adaptor Proteins, Signal Transducing; Adolescent; Adult; Case-Control Studies; Cell Differentiation; Cell Proliferation; Cyclin E; Cyclin-Dependent Kinase 4; DNA-Binding Proteins; Female; Gene Products, env; Gestational Age; Humans; Infant, Newborn; Male; Nuclear Proteins; Oncogene Proteins; Phosphorylation; Placenta; Pre-Eclampsia; Pregnancy; Pregnancy Proteins; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; rhoA GTP-Binding Protein; RNA, Messenger; Signal Transduction; Smad2 Protein; Smad7 Protein; Transcription Factors; Transforming Growth Factor beta; Trophoblasts; Ubiquitin-Protein Ligases; Young Adult

Preeclampsia (PE) is a leading cause of maternal mortality worldwide. Several studies have detected some differentially expressed microRNAs in the preeclamptic placenta, but few of the identified microRNAs demonstrated consistent findings among different research studies. In this study, high-throughput microRNA sequencing (HTS) of 9 preeclamptic and 9 normal placentas was performed. Seventeen microRNAs were identified to be up-regulated, and 8 down-regulated in preeclamptic placentas. Eight differentially expressed microRNAs except one identified in our study were determined to be consistent with at least one previous study, while sixteen were newly found. We performed qRT-PCR with independent 22 preeclamptic placentas and 20 control placentas to verify the differentially expressed microRNAs, and ten microRNAs were validated. The predicted target genes of the aberrantly expressed miR-193b-3p were enriched in the following gene ontology categories: cell motility and migration, cell proliferation and angiogenesis. We also found that miR-193b-3p significantly decreased the migration and invasion of trophoblast (HTR-8/SVneo) cells and that miR-193b-3p could regulate trophoblasts migration and invasion through binding onto the 3'UTR target site of TGF-β2. In conclusion, we identified a list of differentially expressed microRNAs in PE placentas by HTS and provided preliminary evidence for the role of miR-193b-3p in the pathogenesis of preeclampsia.

Topics: Base Sequence; Binding Sites; Cell Movement; Cell Proliferation; Cluster Analysis; Computational Biology; Female; Gene Expression Profiling; Gene Expression Regulation; Gene Ontology; Humans; MicroRNAs; Placenta; Pre-Eclampsia; Pregnancy; Reproducibility of Results; RNA Interference; Signal Transduction; Transforming Growth Factor beta; Transforming Growth Factor beta2

Preeclampsia (PE) is a pregnancy disorder characterized by high blood pressure and proteinuria that can cause adverse health effects in both mother and fetus. There is no current cure for PE other than delivery of the fetus/placenta. While the etiology is unknown, poor placentation due to aberrant signaling of growth and angiogenic factors has been postulated as a causal factor of PE. In addition, environmental contaminants, such as the metal cadmium (Cd), have been linked to placental toxicity and increased risk of developing PE. Here, we use a translational study design to investigate genomic and epigenomic alterations in both placentas and placental trophoblasts, focused on the angiogenesis-associated transforming growth factor-beta (TGF-β) pathway. Genes within the TGF-β pathway displayed increased expression in both the preeclamptic placenta and Cd-treated trophoblasts. In addition, miRNAs that target the TGF-β pathway were also significantly altered within the preeclamptic placenta and Cd-treated trophoblasts. Integrative analysis resulted in the identification of a subset of Cd-responsive miRNAs, including miR-26a and miR-155, common to preeclamptic placentas and Cd-treated trophoblasts. These miRNAs have previously been linked to PE and are predicted to regulate members of the TGF-β pathway. Results from this study provide future targets for PE treatment.

Topics: Cadmium; Case-Control Studies; Environment; Epigenomics; Female; Humans; MicroRNAs; Placenta; Pre-Eclampsia; Pregnancy; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Transforming Growth Factor beta; Trophoblasts

NKp44 and NKp30 splice variant profiles have been shown to promote diverse cellular functions. Moreover, microenvironment factors such as TGF-β, IL-15 and IL-18 are able to influence both NKp44 and NKp30 splice variant profiles, leading to cytokine-associated profiles. Placenta and cancerous tissues have many similarities; both are immunologically privileged sites and both share immune tolerance mechanisms to support tissue development. Therefore, we studied the profiles of NKp44 and NKp30 splice variants in these states by comparing (i) decidua from pregnancy disorder and healthy gestation and (ii) matched normal and cancer tissue. Decidua samples had high incidence of both NKp44 and NKp30. In cancerous state it was different; while NKp30 expression was evident in most cancerous and matched normal tissues, NKp44 incidence was lower and was mostly associated with the cancerous tissues. A NKp44-1dominant inhibitory profile predominated in healthy pregnancy gestation. Interestingly, the NKp44-2/3 activation profile becomes the leading profile in spontaneous abortions, whereas balanced NKp44 profiles were observed in preeclampsia. In contrast, a clear preference for the NKp30a/b profile was evident in the 1st trimester decidua, yet no significant differences were observed for NKp30 profiles between healthy gestation and spontaneous abortions/preeclampsia. Both cancerous and matched normal tissues manifested balanced NKp30c inhibitory and NKp30a/b activation profiles with a NKp44-1dominant profile. However, a shift in NKp30 profiles between matched normal and cancer tissue was observed in half of the cases. To summarize, NKp44 and NKp30 splice variants profiles are tissue/condition specific and demonstrate similarity between placenta and cancerous tissues.

Topics: Abortion, Spontaneous; Decidua; Female; Flow Cytometry; Humans; Immune Privilege; Interleukin-15; Interleukin-18; Killer Cells, Natural; Lymphocytes, Tumor-Infiltrating; Natural Cytotoxicity Triggering Receptor 2; Natural Cytotoxicity Triggering Receptor 3; Neoplasms; Pre-Eclampsia; Pregnancy; RNA Splicing; Transforming Growth Factor beta; Tumor Microenvironment

Circulating immune markers may be associated with preeclampsia but further investigations in early pregnancy and among preeclampsia subtypes are warranted. We examined immune markers in 208 preeclamptic women and 411 normotensive controls.. Our study was nested within the Collaborative Perinatal Project. A total of 242 women had first trimester serum samples and 392 had second trimester serum samples. Preeclampsia was defined as hypertension >20weeks of gestation with proteinuria or pulmonary edema, oliguria, or convulsions. Preterm preeclampsia was defined as preeclampsia with delivery less than 37weeks of gestation. Associations between immune markers RANTES, interleukin (IL)-6, IL4, IL5, IL12, IL10, IL8, IL1-beta, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha and beta, transforming growth factor (TGF)-beta and preeclampsia were explored using a modified version of cox regression developed to address data with non-detectable levels. Models were adjusted for body mass index, gestational age of blood sampling, fetal sex, smoking, socioeconomic status and maternal age.. In first trimester samples, IL-12 was associated with preeclampsia (p=0.0255). IFN-gamma (p=0.0063), IL1-beta (p=0.0006), IL5 (p=0.0422) and TNFr (p=0.0460) were associated with preterm preeclampsia only. In second trimester samples, IL1-beta was associated with preeclampsia (p=0.0180) and term preeclampsia (p=0.0454). After correction for multiple comparisons, only IL1-beta remained associated with preterm preeclampsia in the first trimester (p=0.0288).. Elevated first trimester IL1-beta appears to be associated with preterm preeclampsia. However, few associations were observed in the second trimester. Systemic immune markers alone may not be useful for preeclampsia prediction.

Topics: Adolescent; Adult; Biomarkers; Case-Control Studies; Chemokine CCL5; Cytokines; Female; Humans; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-1beta; Interleukin-4; Interleukin-5; Interleukin-6; Interleukin-8; Lymphotoxin-alpha; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, First; Pregnancy Trimester, Second; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Young Adult

Abnormal levels of High temperature requirement A1 (HtrA1) protein have been repeatedly observed in sera and placentas of preeclampsia patients. To understand the functions of HtrA1 in placentation and in the etiology of preeclampsia, we established HtrA1(-/-) mice. HtrA1(-/-) mice show intrauterine growth retardation, and their placentas are small due to a reduced size of the junctional zone and aberrant vascularization in the labyrinth at the mid-gestation stage. HtrA1 is expressed by Tpbpa-positive trophoblast precursors in the outer ectoplacental cone and junctional zone from embryonic day 7.5 to 10.5. In the HtrA1(-/-) placenta, Tpbpa-positive cell precursors are decreased in the early stage. Spongiotrophoblasts and glycogen trophoblast cells, both of which differentiate from Tpbpa-positive precursors, are consequently decreased in the junctional zone. Fewer spiral artery-associated trophoblast giant cells, another cell type derived from Tpbpa-positive precursors, invade the decidua and associate with maternal arteries in the HtrA1(-/-) placenta than in the wild type placenta. Maternal arteries in the HtrA1(-/-) decidua have narrower lumens, thicker arterial walls, and more vascular smooth muscle cells remaining in the walls than those in the wild type decidua, indicating impaired remodeling of maternal arteries. These results indicate that HtrA1 plays important roles in the differentiation of trophoblasts from Tpbpa-positive precursors in the ectoplacental cone. Insufficient levels of HtrA1 cause poor placental development and intrauterine growth retardation, due to aberrant trophoblast differentiation and consequent defects in maternal artery remodeling, and may contribute to the onset of preeclampsia.

Topics: Animals; Cell Differentiation; Cell Lineage; Decidua; Disease Models, Animal; Female; Fibroblast Growth Factors; High-Temperature Requirement A Serine Peptidase 1; Mice; Mice, Knockout; Phenotype; Placentation; Pre-Eclampsia; Pregnancy; Serine Endopeptidases; Time Factors; Transforming Growth Factor beta; Trophoblasts

Preeclampsia is a potentially fatal pregnancy disorder affecting millions of women around the globe. Dysregulation in gene and protein expression within key biological pathways controlling angiogenesis has been implicated in the development of preeclampsia. Altered CpG methylation, a type of epimutation, may underlie this pathway dysregulation. In the present study, placental tissue from preeclamptic cases and normotensive controls was analyzed for genome-wide differential CpG methylation and concomitant changes in gene expression. A set of 123 genes, representing 19.9% of all genes with altered CpG methylation, was associated with functional changes in transcript levels. Underscoring the complex relationships between CpG methylation and gene expression, here hypermethylation was never associated with gene silencing, nor was hypomethylation always associated with gene activation. Moreover, the genomic region of the CpG mark was important in predicting the relationship between CpG methylation and gene expression. The 123 genes were enriched for their involvement in the transforming growth factor beta (TGF-β) signaling pathway, a known regulator of placental trophoblast invasion and migration. This is the first study to identify CpG hypomethylation as an activator of TGF-β-associated gene expression in the preeclamptic placenta. The results suggest functional epimutations are associated with preeclampsia disease status and the identified genes may represent novel biomarkers of disease.

Topics: Adult; Case-Control Studies; Cluster Analysis; Cohort Studies; Computational Biology; CpG Islands; DNA Methylation; Epigenesis, Genetic; Epigenomics; Female; Gene Expression Profiling; Gene Expression Regulation; Gestational Age; Humans; Placenta; Pre-Eclampsia; Pregnancy; Protein Interaction Maps; Signal Transduction; Transforming Growth Factor beta

Abstract Objective: To research the hypothesis of preeclampsia (PE) is associated with increased systemic inflammatory responses of Th1-type as well as decreased Th2-type responses; we evaluated the maternal plasma levels of IFN-gamma, TNF-alpha, TGF-beta, IL-4, IL-6, IL-10, IL-17, IL-35 and SOCS3 in preeclamptic and healthy pregnants.. This study was conducted with 40 preeclamptic (study group) and 40 normotensive pregnant (control) women in third trimester when they were admitted to the labor and delivery unit. The extracted maternal plasma samples were assayed by an enzyme-linked immunosorbent assay. Statistical analysis was performed by SPSS 16.0 version.. While IFN-gamma and TGF-beta levels of preeclamptic women were significantly higher (p < 0.01), IL-35 and IL-17 levels of preeclamptic women were significantly lower (p < 0.01) than those of controls. The ratios of IFN-gamma/IL-10, IFN-gamma/IL-6, IFN-gamma/IL-4 were significantly high and ratio of IL-35/IL-17 was significantly low in the PE group compared to those in the control group. Maternal plasma SOCS3 levels showed negative correlation with blood pressure and proteinuria severity, but none of the cytokines showed influence on blood pressure and proteinuria after adjusting for maternal and gestational age.. Increased IFN-gamma/TGF-beta production and reduced IL-35/IL-17/SOCS3 production in preeclamptic women may lead to less cytokine inhibitory activity in PE, which may account for the increased proteinuria and blood pressure in PE.

Topics: Adult; Biomarkers; Case-Control Studies; Female; Humans; Interferon-gamma; Interleukin-17; Interleukins; Pre-Eclampsia; Pregnancy; Suppressor of Cytokine Signaling 3 Protein; Suppressor of Cytokine Signaling Proteins; Transforming Growth Factor beta

The "two-stage disorder" theory that is assumed for the etiology of preeclampsia hypothesizes that antiangiogenic and angiogenic factors and/or placental debris play an important role in this disorder. The physiological actions of placental debris occur via the balance between antiangiogenic and angiogenic factors. Accordingly, this balance between antiangiogenic and angiogenic factors should be investigated to elucidate the various pathological features of preeclampsia. Their accurate evaluation is needed to investigate not only antiangiogenic factors (such as sFlt-1 and sEng) and angiogenic factors (such as vascular endothelial growth factor, placental growth factor and transforming growth factor-β) but also the expression level of their receptors such as Flt-1 and Eng. However, it is ethically and technically difficult to investigate the above-mentioned factors at antepartum in human patients. The examination of the ratios of sFlt-1/vascular endothelial growth factor receptor ligands and sEng/transforming vascular endothelial growth factor-β and the use of experimental animal models may help in elucidating various unresolved issues in preeclampsia.

Topics: Angiogenesis Inducing Agents; Angiogenesis Inhibitors; Antigens, CD; Endoglin; Female; Humans; Neovascularization, Pathologic; Patient Acuity; Placenta; Placenta Growth Factor; Pre-Eclampsia; Pregnancy; Pregnancy Proteins; Receptors, Cell Surface; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-1

Topics: Female; Gene Expression Regulation; Humans; MicroRNAs; Nodal Protein; Pre-Eclampsia; Pregnancy; RNA; Transforming Growth Factor beta; Trophoblasts

Preeclampsia is a major disorder of pregnancy and a leading cause of maternal and perinatal morbidity and mortality. MicroRNAs are small noncoding RNAs that regulate gene expression posttranscriptionally. In this study, we examined the expression of miR-376c and found that miR-376c levels were downregulated in both placental and plasma samples collected from preeclamptic patients, when compared with the normal pregnant women at the same gestational stage. Overexpression of miR-376c induced trophoblast cell proliferation, migration, and invasion in HTR8/SVneo cells and promoted placental explant outgrowth. In contrast, inhibition of endogenous miR-376c resulted in a decrease in trophoblast cell invasion and placental explant outgrowth. We identified activin receptor-like kinase 5 (ALK5), a type I receptor for transforming growth factor-β, and ALK7, a type I receptor for Nodal, as targets of miR-376c. Overexpression of miR-376c repressed transforming growth factor-β and Nodal functions, whereas overexpression of ALK5 and ALK7 reversed the effects of miR-376c. These results demonstrate that miR-376c inhibits both ALK5 and ALK7 expression to impair transforming growth factor-β/Nodal signaling, leading to increases in cell proliferation and invasion. An unbalanced Nodal/transforming growth factor-β and miR-376c expression may lead to the development of preeclampsia.

Topics: Cell Proliferation; Female; Gene Expression Regulation; Humans; MicroRNAs; Nodal Protein; Placenta; Pre-Eclampsia; Pregnancy; RNA; Signal Transduction; Transforming Growth Factor beta; Trophoblasts

Endothelial dysfunction is a hallmark of preeclampsia. Desensitization of the phosphoinositide 3-kinase (PI3K)/Akt pathway underlies endothelial dysfunction and haeme oxygenase-1 (HO-1) is decreased in preeclampsia. To identify therapeutic targets, we sought to assess whether these two regulators act to suppress soluble endoglin (sEng), an antagonist of transforming growth factor-β (TGF-β) signalling, which is known to be elevated in preeclampsia.. Vascular endothelial growth factor-A (VEGF-A), fibroblast growth factor (FGF-2), angiopoietin-1 (Ang-1), and insulin, which all activate the PI3K/Akt pathway, inhibited the release of sEng from endothelial cells. Inhibition of the PI3K/Akt pathway, by overexpression of phosphatase and tensin homolog (PTEN) or a dominant-negative isoform of Akt (Akt(dn)) induced sEng release from endothelial cells and prevented the inhibitory effect of VEGF-A. Conversely, overexpression of a constitutively active Akt (Akt(myr)) inhibited PTEN and cytokine-induced sEng release. Systemic delivery of Akt(myr) to mice significantly reduced circulating sEng, whereas Akt(dn) promoted sEng release. Phosphorylation of Akt was reduced in preeclamptic placenta and this correlated with the elevated level of circulating sEng. Knock-down of Akt using siRNA prevented HO-1-mediated inhibition of sEng release and reduced HO-1 expression. Furthermore, HO-1 null mice have reduced phosphorylated Akt in their organs and overexpression of Akt(myr) failed to suppress the elevated levels of sEng detected in HO-1 null mice, indicating that HO-1 is required for the Akt-mediated inhibition of sEng.. The loss of PI3K/Akt and/or HO-1 activity promotes sEng release and positive manipulation of these pathways offers a strategy to circumvent endothelial dysfunction.

Topics: Angiopoietin-1; Animals; Antigens, CD; Endoglin; Endothelium, Vascular; Female; Fibroblast Growth Factor 2; Heme Oxygenase-1; Humans; Insulin; Mice; Mice, Inbred C57BL; Phosphatidylinositol 3-Kinases; Phosphorylation; Pre-Eclampsia; Pregnancy; Proto-Oncogene Proteins c-akt; PTEN Phosphohydrolase; Receptors, Cell Surface; RNA, Messenger; Signal Transduction; Transforming Growth Factor beta; Umbilical Veins; Vascular Endothelial Growth Factor A

Emerging evidence has shown that podocyte injury and reduced specific podocyte protein expressions contribute to proteinuria in preeclampsia. We collected urine specimens from women with preeclampsia to study whether podocyte-specific protein shedding is associated with renal barrier dysfunction. Urine specimens from women with normal pregnancies and from pregnant women complicated by chronic hypertension were used for comparison. We determined soluble podocyte slit protein nephrin levels in the urine specimens. Podocalyxin, βig-h3, and VEGF concentrations were also measured. We found that nephrin and podocalyxin were barely detectable in the urine specimens from normal pregnant women and from women with chronic hypertension. In preeclampsia, urinary nephrin and podocalyxin concentrations were significantly increased and highly correlated to each other, r(2) = 0.595. Nephrin and podocalyxin were also correlated with urine protein concentrations. βig-h3 was detected in the urine specimens from women with preeclampsia, and it is highly correlated with nephrin and podocalyxin concentrations in preeclampsia. βig-h3 was undetectable in normal pregnancy and pregnancy complicated by chronic hypertension. Elevated VEGF levels were also found in women with preeclampsia compared with those of normal pregnancy and pregnancy complicated by chronic hypertension. These results provide strong evidence that podocyte protein shedding occurs in preeclampsia, and their levels are associated with proteinuria. The finding of urinary βig-h3 excretion in preeclampsia suggests that increased transforming growth factor activity might also be involved in the kidney lesion in this pregnancy disorder.

Topics: Adult; Biomarkers; Case-Control Studies; Extracellular Matrix Proteins; Female; Humans; Hypertension; Kidney; Membrane Proteins; Pre-Eclampsia; Pregnancy; Pregnancy Complications, Cardiovascular; Sialoglycoproteins; Signal Transduction; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

Topics: Extracellular Matrix Proteins; Female; Humans; Membrane Proteins; Pre-Eclampsia; Pregnancy; Pregnancy Complications, Cardiovascular; Sialoglycoproteins; Transforming Growth Factor beta

Pre-eclampsia is a disorder that results in significant feto-maternal complications with yet no definitive pharmacologic intervention. One postulated etiologic mechanism is an imbalance between circulating pro-angiogenic and anti-angiogenic factors. We investigated these factors sequentially throughout pregnancy (19-21 days) in our rat model of pre-eclampsia, which involves the imposition of excessive volume expansion.. We evaluated the status of the pro-angiogenic and anti-angiogenic factors at the following time points: 3-5, 7-10 and 17-20 days of gestation.. We have previously determined that the urinary excretion of the circulating bufodienolide, marinobufagenin, is elevated at the 3- to 5-day time period, prior to the advent of hypertension and proteinuria. At 3-5 days of pregnancy, there was no evidence of angiogenic imbalance in the normal pregnant (NP) and 'pre-eclamptic' (PDS) rats. At the 7- to 10-day time point, plasma PlGF was greater in the NP rats than in the PDS group (p < 0.05). The plasma sFlt-1/PlGF ratio in the PDS animals was greater than that in the NP rats (p < 0.05). The placental sFlt-1 and sFlt-1/PlGF ratio were greater in the PDS rats than in NP rats (p < 0.05). These changes were also present at the 17- to 20-day time point in both plasma and placenta. The administration of resibufogenin, an antagonist of marinobufagenin, early in pregnancy, prevented angiogenic imbalance.. We conclude that angiogenic imbalance plays a role in the pathogenesis of pre-eclampsia in this rat model. Furthermore, the earliest event in the pathogenetic sequence appears to be the secretion and elaboration of marinobufagenin.

Topics: Analysis of Variance; Angiogenic Proteins; Animals; Blood Pressure; Bufanolides; Creatinine; Endoglin; Female; Gestational Age; Hematocrit; Intracellular Signaling Peptides and Proteins; Models, Animal; Placenta Growth Factor; Pre-Eclampsia; Pregnancy; Pregnancy Proteins; Proteinuria; Rats; Rats, Sprague-Dawley; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-1

Topics: Angiogenesis Inducing Agents; Clinical Trials as Topic; Female; HELLP Syndrome; Humans; Kidney Diseases; Placenta Growth Factor; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Pregnancy Proteins; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-1

The objective of this investigation was to characterize soluble endoglin (sEng) concentrations in second-trimester serum of women who either develop preeclampsia or have a normal pregnancy.. Single second-trimester serum samples obtained from healthy, nonsmoking women who subsequently developed severe preeclampsia (n = 48) or from healthy nonsmoking women who experienced a normal pregnancy (n = 56) were measured by enzyme-linked immunosorbent assay. Data were reported as mean +/- standard deviation.. Maternal age or gestational age at time of sample was not different between the 2 groups. Patients who later developed preeclampsia delivered earlier, had smaller infants, and had a higher mean arterial pressure than controls. Patients who later developed severe preeclampsia had elevated sEng, compared with those with normal pregnancy (6.19 +/- 2.1 vs 5.00 +/- 1.0 ng/mL, P = .02).. Soluble endoglin is elevated in second-trimester maternal serum in patients destined to develop severe preeclampsia.

Topics: Adult; Antigens, CD; Biomarkers; Case-Control Studies; Endoglin; Female; Gestational Age; Humans; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Second; Receptors, Cell Surface; Signal Transduction; Transforming Growth Factor beta

Preeclampsia is a pregnancy-specific hypertensive syndrome that causes substantial maternal and fetal morbidity and mortality. Maternal endothelial dysfunction mediated by excess placenta-derived soluble VEGF receptor 1 (sVEGFR1 or sFlt1) is emerging as a prominent component in disease pathogenesis. We report a novel placenta-derived soluble TGF-beta coreceptor, endoglin (sEng), which is elevated in the sera of preeclamptic individuals, correlates with disease severity and falls after delivery. sEng inhibits formation of capillary tubes in vitro and induces vascular permeability and hypertension in vivo. Its effects in pregnant rats are amplified by coadministration of sFlt1, leading to severe preeclampsia including the HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome and restriction of fetal growth. sEng impairs binding of TGF-beta1 to its receptors and downstream signaling including effects on activation of eNOS and vasodilation, suggesting that sEng leads to dysregulated TGF-beta signaling in the vasculature. Our results suggest that sEng may act in concert with sFlt1 to induce severe preeclampsia.

Topics: Adult; Amino Acid Sequence; Animals; Antigens, CD; Endoglin; Endothelial Cells; Female; Gestational Age; Hemodynamics; Humans; Kidney; Liver; Mice; Mice, Knockout; Middle Aged; Molecular Sequence Data; Nitric Oxide Synthase Type III; Placenta; Pre-Eclampsia; Pregnancy; Pregnancy, Animal; Rats; Rats, Sprague-Dawley; Receptors, Cell Surface; Signal Transduction; Transforming Growth Factor beta; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor Receptor-1

To study the change and significance of the expression of transforming growth factor-beta 1 (TGF-beta1), vascular cell adhesion molecule-1 (VCAM-1) and endothelium-selectin (E-selectin) in placenta of patients with pre-eclampsia.. Twenty normal pregnant women (control group) and 40 women with pre-eclampsia (pre-eclampsia group, including 16 women with mild pre-eclampsia and 24 women with severe pre-eclampsia) were selected. The cellular distribution of TGF-beta1, VCAM-1 and E-selectin in placenta in both groups was determined by immunohistochemistry, and the mean density was measured by computer image analysis system.. (1) The level of TGF-beta1 in placental villous syncytiotrophoblast of pre-eclampsia group (70.7 +/- 0.5) was significantly higher than that of control group (70.3 +/- 0.6), while the level of VCAM-1 and E-selectin in pre-eclampsia group (VCAM-1: 82.5 +/- 0.5, E-selectin: 53.5 +/- 0.5) was significantly lower than that of control group (VCAM-1: 82.8 +/- 0.3, E-selectin: 53.8 +/- 0.4) (P < 0.05). However, there were no significant differences in women with mild pre-eclampsia (TGF-beta1: 70.6 +/- 0.6, VCAM-1: 82.4 +/- 0.6, E-selectin: 53.4 +/- 0.5) and severe ones (TGF-beta1: 70.8 +/- 0.4, VCAM-1: 82.6 +/- 0.5, E-selectin: 53.6 +/- 0.5) (P > 0.05); (2) The level of E-selectin in placental villous capillary endothelial cells of pre-eclampsia group (63.0 +/- 0.5) was significantly higher than that of control group (62.6 +/- 0.4) (P < 0.05), while there was no significant difference in women with mild pre-eclampsia (63.2 +/- 0.4) and severe pre-eclampsia (62.9 +/- 0.5) (P > 0.05).. TGF-beta1, VCAM-1 and E-selectin are not only related to placenta shallow bed of pre-eclampsia, but also participate in pathogenic process of vascular endothelial damage of pre-eclampsia.

Topics: Adult; E-Selectin; Female; Humans; Immunohistochemistry; Placenta; Pre-Eclampsia; Pregnancy; Transforming Growth Factor beta; Trophoblasts; Vascular Cell Adhesion Molecule-1

PL74, a novel member of the TGFbeta superfamily that has highest expression in placenta, is a multifunctional peptide that can induce differentiation, inhibit inflammatory stimulation of TNFalpha, and execute apoptosis after p53 overexpression and cytotoxic injury. To study its expression and function in placenta and preeclampsia, we first determined mRNA expression in nine normal and 10 preeclamptic placentas. PL74 mRNA was overexpressed by 57.3% in preeclampsia. Transfection of PL74 into term cytotrophoblasts resulted in increased apoptosis by terminal uridine deoxynucleotidyl nick end labeling labeling (control, 2.8 +/- 0.5%; PL74, 19.1 +/- 0.2%; P < 0.005). Addition of PL74 protein to HTR8/SVneo extravillous cytotrophoblast cells showed a dose-response (0-100 ng/ml) inhibition of [3H]thymidine uptake and increase in apoptosis shown by terminal uridine deoxynucleotidyl nick end labeling and histone-associated DNA fragment ELISA (control, 0.11 +/- 0.01 absorbance units; PL74, 0.21 +/- 0.01; P < 0.01). PL74 did not alter cytotrophoblast invasion using a Matrigel in vitro invasion assay. Cytokine regulation of PL74 mRNA expression in term cytotrophoblasts showed that epidermal growth factor and IFNgamma increased PL74 expression, but TGFbeta and TNFalpha had no effect. Transfection of antisense PL74 into term cytotrophoblast cells resulted in an inhibition of spontaneous differentiation at 2 and 24 h of culture (control vector, 30.8 +/- 3.1% and 26.4 +/- 1.2%; antisense PL74, 17.6 +/- 1.8%and 12.6 +/- 1.4% syncytial units, at 2 and 24 h respectively; P < 0.01). We conclude that PL74 is overexpressed in preeclampsia and may thus promote apoptosis of cytotrophoblasts at the expense of differentiation. PL74 secretion is induced by IFNgamma and may play a role in abnormal placental responses in preeclampsia.

Topics: Apoptosis; Biopsy; Cell Differentiation; Cells, Cultured; Female; Humans; Interferon-gamma; Placenta; Pre-Eclampsia; Pregnancy; RNA, Messenger; Transforming Growth Factor beta; Trophoblasts; Tumor Necrosis Factor-alpha

To investigate feto-maternal bone turnover in normal pregnancy and preeclampsia and to test the hypothesis whether the reported low bone mass at birth in small-for-gestational age infants is associated with decreased bone formation or increased bone resorption.. Thirty-two patients with preeclampsia (17 mild and 15 severe) and 20 normotensive women (controls) with singleton gestations in the third trimester participated in this study. Furthermore, 25 nonpregnant healthy women were chosen as nonpregnant controls. Maternal 24-hour urine specimens and venous blood samples were collected. In addition, fetal cord blood and the first voided neonatal urine were also collected. The freshly separated sera were assayed for osteocalcin (OC) and carboxy-terminal propeptide of type 1 collagen (PICP) by radioimmunoassay. Urine samples were assayed for N-telopeptide of type 1 collagen (NTx) by enzyme-linked immunosorbent assay.. Maternal and cord serum OC and PICP levels were significantly decreased in severe preeclampsia, whereas maternal and first-voided neonatal urinary NTx level were significantly increased compared to the corresponding levels of controls. In both mother and fetus, the coupling index of markers of bone turnover in normal pregnancy or mild preeclampsia was in favor of bone formation, whereas in severe preeclampsia the markers suggested marked bone resorption.. Increased bone resorption and decreased bone formation occur in preeclampsia in both mother and fetus, being more pronounced in the latter. The increased osteoclastic activity in preeclampsia may be attributed to increased RANKL induced by increased interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and transforming growth factor beta2 (TGF-beta2) production.

Topics: Adult; Bone Resorption; Carrier Proteins; Case-Control Studies; Collagen; Collagen Type I; Enzyme-Linked Immunosorbent Assay; Female; Fetal Blood; Humans; Infant, Newborn; Infant, Small for Gestational Age; Interleukin-6; Maternal-Fetal Exchange; Membrane Glycoproteins; Osteocalcin; Osteoclasts; Peptides; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Third; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Transforming Growth Factor beta; Transforming Growth Factor beta2; Tumor Necrosis Factor-alpha

We examined the relationship between maternal plasma transforming growth factor-beta1 (TGF-beta1) concentrations and risk of preeclampsia among women delivering at Harare Maternity Hospital in Zimbabwe. We evaluated the relationship in the context of maternal systemic inflammation using plasma tumor necrosis factor-a soluble receptor p55 (sTNFp55) as a marker.. 132 women with preeclampsia and 180 controls were included in this case-control study analysis. Maternal post-diagnosis plasma TGF-beta1 and sTNFp55 concentrations were determined using immunoassays. Logistic regression procedures were used to estimate odds ratios (OR) and 95% confidence intervals (CI) adjusted for confounders.. A linear increase in preeclampsia risk was observed with increasing quartiles of TGF-beta1 concentrations (p<0.01). Women whose TGF-beta1 concentrations were >or=25.1 ng/ml (quartile 4) had a 2.5-fold (95% CI 1.2-5.6) increased risk of preeclampsia as compared with those women whose concentrations were <11.2 ng/ml (quartile 1). Relative to women with no evidence of systemic inflammation and no elevated TGF-beta1 concentrations, those women who were jointly positive for elevated TGF-beta1 and sTNFp55 concentrations experienced a 5.3-fold (95% CI 2.3-12.0) increased risk of preeclampsia.. Overall, we noted that elevated TGF-beta1 is associated with an increased risk of preeclampsia. We also noted that the preeclampsia risk is exaggerated in the presence of maternal systemic inflammation.

Topics: Adult; Biological Factors; Case-Control Studies; Female; Humans; Pre-Eclampsia; Pregnancy; Prenatal Diagnosis; Receptors, Tumor Necrosis Factor, Type I; Transforming Growth Factor beta; Transforming Growth Factor beta1; Zimbabwe

An understanding of how the fetus escapes the maternal immune system may be relevant for the prevention of transplant rejection. There is evidence that the same immunosuppressive cytokines contribute to a successful pregnancy and transplant success. Transforming growth factor beta (TGF-beta) is a multifunctional cytokine that exhibits potent immunoregulatory and anti-inflammatory properties and may prolong graft survival. Recent reports suggest a role for TGF-beta in the generation of T-regulatory lymphocytes. Also, the role of TGF-beta in trophoblast differentiation and hypertension prompted us to evaluate maternal serum TGF-beta1 levels in normal allopregnant women and in pregnancies complicated by preeclampsia (PE), a disorder characterized by increased blood pressure, proteinuria, and end organ damage. Sixty-one pregnant preeclamptic women (32 cases with severe and 29 with mild PE), 22 normotensive healthy pregnant, and 20 nonpregnant controls formed the study groups. The active form of serum TGF-beta1 was investigated by an indirect ELISA technique. The results showed that TGF-beta1 was highly expressed in all three pregnant groups compared with the nonpregnant controls. No changes in TGF-beta1 serum levels was found in PE compared with a normal pregnancy. The results suggest that: (1) TGF-beta1 may function as a regulatory factor in fetal allograft survival during pregnancy and (2) TGF-beta1 does not have a pathophysiological role in PE.

Topics: Adult; Female; Fetal Tissue Transplantation; Graft Survival; Humans; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Third; Reference Values; Transforming Growth Factor beta; Transforming Growth Factor beta1; Transplantation, Homologous

In a case-control study of 100 preeclamptics and 100 controls, we assessed plasma transforming growth factor-beta1 (TGF-beta1) concentrations in relation to preeclampsia risk among Peruvian women with and without systemic inflammation.. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs).. The OR of preeclampsia increased across quartiles of TGF-beta1 concentrations. Women with elevated TGF-beta1 and a proinflammatory profile experienced the highest risk of preeclampsia (OR = 15.4, 95% CI 4.7-50.4).. Our results confirm an association between TGF-beta1 and risk of preeclampsia and extend the literature by indicating a strong association in women with systemic inflammation.

Topics: Adolescent; Adult; Antigens, CD; Biomarkers; Case-Control Studies; Cytokines; Female; Humans; Inflammation Mediators; Maternal Age; Maternal Welfare; Peru; Pre-Eclampsia; Pregnancy; Receptors, Tumor Necrosis Factor; Receptors, Tumor Necrosis Factor, Type I; Risk Factors; Statistics as Topic; Transforming Growth Factor beta; Transforming Growth Factor beta1

Normal placental development is dependent on the orchestrated differentiation of cytotrophoblast (CTB) cells. This study was aimed at studying cytotrophoblast cells from normal and preeclamptic pregnancies in a three-dimensional spheroid-based cell culture model. First trimester cytotrophoblast cells cultured as spheroids maintain their high proliferative and invasive phenotype and respond to different cytokines upon stimulation in a three-dimensional invasion assay. In contrast, third trimester cytotrophoblast spheroids maintain their quiescent nonproliferating phenotype and invasion can only be induced by EGF. Contrasting the regular spheroidal arrangement of cytotrophoblast cells from normal third trimester pregnancies, spheroidal organization of preeclamptic cytotrophoblast cells is disturbed and the cells downregulate CD105 in vivo and in vitro. Furthermore, the invasion of both normal and preeclamptic third trimester, but not first trimester cytotrophoblast cells, is inhibited by pro-inflammatory cytokines. Plasma samples from pregnant women with preeclampsia significantly stimulate the invasion of first trimester cytotrophoblast cells and the sprouting of human umbilical vein endothelial cells (HUVECs) compared to plasma samples from healthy pregnant women. Taken together, the data establish the spheroidal cytotrophoblast model as a powerful system to mimic the in vivo phenotype of first and third trimester and preeclamptic cytotrophoblast cells and demonstrate that plasma-derived factors modulate the differentiation of cytotrophoblast cells.

Topics: Angiopoietin-2; Antigens, CD; Cell Differentiation; Cell Division; Cells, Cultured; Cytokines; Down-Regulation; Epidermal Growth Factor; Female; Fibroblast Growth Factor 2; Humans; Phenotype; Placenta Growth Factor; Pre-Eclampsia; Pregnancy; Pregnancy Proteins; Pregnancy Trimester, First; Pregnancy Trimester, Third; Spheroids, Cellular; Transforming Growth Factor beta; Transforming Growth Factor beta1; Trophoblasts; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Excessive Th1 activity in peripheral blood plays a probable role in the pathogenesis of preeclampsia. The aim of the study was to investigate whether disturbed local immune reactions are also present in decidua.. Flow cytometric analysis of CD3, CD19, CD56/CD16, CD4, CD8, CD4/CD29, CD4/CD45RA, CD4/CD45RO, CD8/CD28, CD3/CD69 lymphocyte subsets isolated from third trimester decidua of pregnants with preeclampsia (n=21) and pregnant controls (n=11) subjected to elective caesarean sections. Spontaneous and phytohemaglutynine stimulated "in vitro" secretion of IL-2, IL-4, IL-6, IL-10, IL-12, IFN-gamma and TGF-beta by decidual lymphocytes was studied by ELISA. For the statistical significance of differences between the groups the U Mann-Whitney test was performed (confidence interval P<0.05).. Preeclamptic patients were characterized with an increased percentage of the CD3-/CD56+CD16+, CD8+/CD28+ and decreased percentage of CD3+, CD19+, CD4+/CD45RA+ lymphocytes. The profile of secreted cytokines shifts in favor of Th1 activity (extremely high IFN-gamma and low IL-6 and IL-10 secretion). Decidual IL-12 secretion in preeclamptic patients is decreased compared to controls.. Changes in NK and T lymphocyte subsets followed with Th1 cytokine IFN-gamma over-activity, could affect local immunoregulatory mechanisms in third trimester decidua of preeclamptic patients.

Topics: Antigens, CD; Cytokines; Decidua; Female; Flow Cytometry; Humans; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-2; Interleukin-4; Interleukin-6; Killer Cells, Natural; Lymphocyte Subsets; Lymphocytes; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Third; Reference Values; Th1 Cells; Transforming Growth Factor beta

To determine the plasma concentrations of placental growth factor (PLGF), vascular endothelial growth factor (VEGF), transforming growth factor-beta1 (TGF-beta1), soluble tumor necrosis factor alpha receptor (sTNFp55), interleukin-2 receptor (IL-2R), and interleukins 6 and 10 (IL-6, IL-10) in normotensive and preeclamptic women, and to evaluate the correlations between these cytokines and the diastolic blood pressure and fibronectin levels.. A prospective case-control study. Thirty-five women with preeclampsia were compared with 34 healthy women with uncomplicated pregnancies. Peripheral venous blood samples were obtained and plasma levels of PLGF, VEGF, TGF-beta1, sTNFp55, IL-2R, IL-6 and IL-10 were measured by an enzyme-linked immunoassay and fibronectin by a radial immundiffusion technic.. In preeclampsia PLGF and VEGF levels were significantly lower, and TGF-beta1, sTNFp55, IL-2R, IL-6 and IL-10 levels were significantly higher than in normotensive pregnancy (p < 0.001). The plasma levels of PLGF and VEGF significantly decreased, whereas TGF-beta1, sTNFp55, IL-2R, IL-6 and IL-10 levels significantly increased with the increments in diastolic blood pressure and fibronectin levels (p < 0.001).. Altered concentrations of various cytokines might explain the shallow placentation and endothelial cell dysfunction described in preeclampsia. The clinical severity of preeclampsia seems to correlate with the severity of the cytokine abnormalities.

Topics: Adult; Blood Pressure; Case-Control Studies; Cytokines; Diastole; Endothelial Growth Factors; Female; Fibronectins; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-6; Interleukin-8; Lymphokines; Placenta Growth Factor; Pre-Eclampsia; Pregnancy; Pregnancy Proteins; Receptors, Interleukin-2; Receptors, Tumor Necrosis Factor; Transforming Growth Factor beta; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

The semiquantitative reverse transcription polymerase chain reaction was employed to detect the expression of transforming growth factor beta (TGF-beta) and insulin-like growth factor (IGF) in complete hydatidiform mole, normal first-trimester villi, the normal term placenta (after vaginal/abdominal deliver) and the preeclamptic placenta at term. The expression of IGF-I mRNA was seen in all five tissues, but its level was much lower in the term placental tissues with preeclampsia than in other tissues. The content of IGF-I mRNA in villous tissues from molar pregnancy was slightly higher than in normal first-trimester villi. IGF-II mRNA was detected at similar levels in all three sorts of term placental tissues. However, the expression level of IGF-II mRNA in tissues of complete molar pregnancy was significantly lower than in normal first-trimester villi. TGF-beta(3) was found expressed in all five tissues, while TGF-beta(1) and TGF-beta(2) mRNA were not detected. Compared to the normal first-trimester villi, the expression of TGF-beta(3) in complete hydatidiform molar tissues was comparatively higher. Furthermore, the expression levels of TGF-beta(3) in the preeclamptic placenta and the normal placenta after cesarean birth were higher than in the placenta after vaginal delivery. We concluded that, the change of TGF-beta and IGF expression in placental tissues might be involved in the development of trophoblastic diseases of pregnancy.

Topics: Adult; Chorionic Villi; Electrophoresis, Agar Gel; Female; Humans; Hydatidiform Mole; Insulin-Like Growth Factor I; Insulin-Like Growth Factor II; Placenta; Pre-Eclampsia; Pregnancy; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta

To investigate the plasma and placental levels of interleukin-10 (IL-10), transforming growth factor-beta1 (TGF-beta1), and epithelial-cadherin (E-cadherin) in normotensive and preeclamptic pregnancies.. The study population consisted of 33 women with normotensive pregnancy and 35 women with preeclampsia. Peripheral venous blood samples were collected before labor (35.3 +/- 1.1 and 34.2 +/- 3.4 weeks' gestation for normotensive and preeclamptic pregnancies, respectively), and placental tissues were obtained after delivery. Maternal plasma and placental homogenate IL-10, TGF-beta1, and E-cadherin levels were determined by enzyme-linked immunosorbent assay.. The mean plasma and placental levels of IL-10, TGF-beta1, and E-cadherin were significantly higher in preeclamptic than normotensive patients (P <.001). The plasma and placental levels of IL-10, TGF-beta1, and E-cadherin significantly increased with the increments in diastolic blood pressure (P <.001).. IL-10, TGF-beta1, and E-cadherin may be involved in the pathologic process of preeclampsia. The pathophysiologic changes associated with preeclampsia may stem in part from the overproduction of these placental mediators.

Topics: Adult; Analysis of Variance; Biomarkers; Cadherins; Case-Control Studies; Cohort Studies; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-10; Placenta; Pre-Eclampsia; Pregnancy; Probability; Reference Values; Regression Analysis; Sensitivity and Specificity; Transforming Growth Factor beta; Transforming Growth Factor beta1

To detect the immunoreactivity of insulin-like growth factor-I, insulin-like growth factor-binding proteins-1 and -3 and transforming growth factor beta-3 in the umbilical cords of normal and preeclamptic patients.. Umbilical cords were obtained from 15 normal and 15 preeclamptic patients. Immunoreactivities were determined using either indirect immunofluorescence or immunoperoxidase techniques on formalin-fixed, paraffin-embedded sections. Staining intensity was graded by a semiquantitative scoring method. The results were compared by Mann-Whitney U-test.. The umbilical cords were thinner and the vessels were hypoplastic in the preeclamptic group. Moderate staining intensity for insulin-like growth factor-I, insulin-like growth factor binding protein-1 and -3 and transforming growth factor-beta 3 was observed in normal patients. The preeclamptic group had mild and strong intensities for insulin-like growth factor-I and insulin-like growth factor binding protein-1, respectively, and intensity for insulin-like growth factor binding protein-3 did not change, but diffuse and increased intensity was observed for transforming growth factor-beta 3.. Changes in the intensity of insulin-like growth factor-I and its major binding protein and the transformation of growth factor-beta 3 may play a role in the pathogenesis of preeclampsia by altering the structure and responsiveness of the umbilical cord.

Topics: Adult; Case-Control Studies; Female; Formaldehyde; Humans; Immunoenzyme Techniques; Immunohistochemistry; Insulin-Like Growth Factor Binding Protein 1; Insulin-Like Growth Factor Binding Protein 3; Insulin-Like Growth Factor I; Paraffin Embedding; Pre-Eclampsia; Pregnancy; Staining and Labeling; Transforming Growth Factor beta; Transforming Growth Factor beta3; Umbilical Cord

The aim of the study was to determine the concentrations of TGF-beta, PDGF and VEGF growth factors in peripheral blood. The examinations were carried out in the group of pregnant women with preeclampsia. The results were compared to those obtained in a control group of healthy pregnant women. The statistically significant increase in all examined growth factors was observed in women with preeclampsia in comparison to healthy women. The results suggest that growth factors may play an important role in etiology and pathophysiology of preeclampsia.

Topics: Adult; Case-Control Studies; Endothelial Growth Factors; Female; Humans; Intercellular Signaling Peptides and Proteins; Lymphokines; Platelet-Derived Growth Factor; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Third; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

1. Pre-eclampsia is a human disease of pregnancy characterized by high blood pressure, proteinuria and end-organ damage, if severe. Pre-eclampsia is thought to be related to changes in early placental development, with the formation of a shallower than normal placental bed. 2. Transforming growth factor (TGF)-beta1 is a multifunctional fibrogenic growth factor involved in immune regulation that is elevated in some populations with a high risk of hypertensive end-organ disease related to increases in endothelin release. Transforming growth factor-beta1 is also an important factor in placental implantation. Alterations in TGF-beta1 may be related to abnormal placental development in early pregnancy and, thus, are a candidate for the development of hypertension in pre-eclampsia. 3. The aim of the present study was to examine the placental distribution and serum concentration of TGF-beta1 in patients with pre-eclampsia compared with normal pregnancy. 4. Patients with pre-eclampsia (n = 12) were compared with patients with normal pregnancy (n = 14). Transforming growth factor-beta1 was determined by TGF-beta1 Max ELISA (Promega, Madsion, WI, USA) after serum dilution (1/150) and acid activation. Placental distribution was determined by immunostaining with TGF-beta1 (Santa Cruz, Santa Cruz, CA, USA; 20 ng/mL) and the villi and decidual trophoblast were scored for intensity and extent of staining. 5. Patients with pre-eclampsia had a mean gestational age of 36 weeks, whereas those with a normal pregnancy had a mean gestational age of 39.0 +/- 0.4 weeks. There was no difference in TGF-beta1 concentration between the two groups (mean (+/-SEM) 27.1 +/- 1.0 vs 26.4 +/- 0.7 pg/mL for normal pregnancy and pre-eclampsia, respectively; P = 0.73, Mann-Whitney U-test). There was no correlation between systolic or diastolic blood pressure and TGF-beta1 concentration (regression analysis P = 0.4 and 0.2). Immunostaining was absent in the villous trophoblast cells and endovascular and extravillous trophoblast of term placentas. 6. Although TGF-beta1 is present in trophoblast cells in early pregnancy during placental development, TGF-beta1 concentrations were not increased in the placenta at term in pre-eclampsia and there was no correlation between blood pressure and serum TGF-beta1, suggesting that TGF-beta1 does not play a role in the development of late gestation pre-eclampsia and hypertension.

Topics: Adult; Chi-Square Distribution; Female; Humans; Hypertension; Placenta; Pre-Eclampsia; Pregnancy; Statistics, Nonparametric; Transforming Growth Factor beta; Transforming Growth Factor beta1

Topics: Abortion, Habitual; Animals; Apoptosis; Complement System Proteins; Corticotropin-Releasing Hormone; Female; Fertilization in Vitro; Fetus; Histocompatibility Antigens Class I; HLA Antigens; HLA-G Antigens; Humans; Immune System; Immune Tolerance; Infertility, Female; Killer Cells, Natural; Male; Mice; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Reproduction; Semen; T-Lymphocytes; Transforming Growth Factor beta; Trophoblasts; Tryptophan Oxygenase; Uterus

Recently it has been suggested that TGF-beta(1) may play an important role in trophoblast invasion of spiral arteries in pre-eclampsia. This study was designed to investigate TGF-beta(1) levels in pre-eclampsia. Platelet depleted plasma TGF-beta(1) levels were measured by ELISA in 43 normotensive healthy non-pregnant women, 30 normotensive healthy pregnant and 42 pre-eclamptic women: TGF-beta(1) levels were increased significantly in the pre-eclamptics (4.9+/-0.9 ng/ml) compared to the normotensive pregnant (1.9+/-0.54 ng/ml; P<0.001) and normotensive non-pregnant groups (2.1+/-0.41 ng/ml; P<0.0001). There was no significant difference in TGF-beta(1) levels between normotensive pregnant and non-pregnant groups (1.9+/-0.54 vs. 2.1+/-0.41 ng/ml; P=0.2). There was a highly significant correlation between neonatal weight (r=0.63; P<0.0001) and TGF-beta(1) levels. The finding of increased levels of plasma TGF-beta(1) suggests an aetiological role of this peptide in pre-eclampsia.

Topics: Adolescent; Adult; Birth Weight; Female; Humans; Pre-Eclampsia; Pregnancy; Transforming Growth Factor beta; Transforming Growth Factor beta1

Isoforms of transforming growth factor-beta (TGF-beta1) are thought to be involved in the pathogenesis of pre-eclampsia. Data with respect to TGF-beta1 are controversial. We examined the correlation between TGF-beta1 serum levels and the occurrence and severity of pre-eclampsia.. Transforming growth factor-beta 1 serum levels were measured using a commercially available enzyme-linked immunosorbent assay in 44 women with pre-eclampsia and 44 healthy pregnant women. Results were correlated with clinical data.. No difference in TGF-beta1 serum levels was assessed between women with pre-eclampsia and healthy pregnant women. Transforming growth factor-beta 1 serum levels were not associated with the severity of the disease and were not correlated with clinical maternal (blood pressure, proteinuria) and fetal (5-min APGAR score, umbilical cord pH values, birth weight) parameters.. Our data support the assumption that in contrast to other isoforms, TGF-beta1, as evidenced by serum TGF-beta1 levels, does not seem to be involved in the pathogenesis of pre-eclampsia.

Topics: Adult; Biomarkers; Case-Control Studies; Cohort Studies; Female; Humans; Pre-Eclampsia; Pregnancy; Probability; Reference Values; Retrospective Studies; Sensitivity and Specificity; Transforming Growth Factor beta; Transforming Growth Factor beta1

Members of the TGF-beta family have been shown to play an important role in numerous tissues during development. In the present study we have investigated the spatial and temporal expression of TGF-beta 3, in human umbilical cord development. Total TGF-beta 3 protein content, assessed by immunoblotting, increased with advancing gestation as did immunostaining and mRNA in Wharton's jelly fibroblasts. Immunohistochemical analysis revealed that TGF-beta 3 was present in all cell types. Temporal changes in TGF-beta 3 expression were observed in the vascular smooth muscle cells, such that with advancing gestation TGF-beta 3 protein expression and became mostly restricted to the extracellular compartment of the vascular media. This was associated with a decrease in TGF-beta 3 mRNA expression in umbilical vascular smooth muscle cells. Of clinical significance, umbilical cords from pregnancies complicated by pre-eclampsia, showed a significant reduction in total TGF-beta 3 protein expression when compared to those of age-matched patients. Both TGF-beta 3 mRNA and protein expression were downregulated in the endothelium and smooth muscle layers of the umbilical arteries, as well as in the Wharton jelly fibroblasts. Our data demonstrate that during umbilical cord development TGF-beta 3 expression is spatially and temporally regulated and that TGF-beta 3 expression is altered in umbilical cords of pregnancies complicated by pre-eclampsia. We speculate that the downregulation of TGF-beta 3 expression found in pre-eclamptic umbilical cord may contribute to the abnormal structure and mechanical properties seen in these pathological umbilical cords.

Topics: Case-Control Studies; Down-Regulation; Female; Gene Expression Regulation, Developmental; Humans; Immunohistochemistry; In Situ Hybridization; Pre-Eclampsia; Pregnancy; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta3; Umbilical Cord

In many pre-eclamptic women the placentation process seems to be disturbed. Our objective was to investigate if disturbed placentation in pre-eclamptic women may be recognized in early second trimester as altered plasma levels of factors involved in the formation of the uteroplacental unit.. In a prospective study of 2190 pregnant women we compared plasma leptin, transforming growth factor-beta(1) (TGF-beta(1)) and plasminogen activator inhibitor type 2 (PAI-2) concentrations at 18 weeks' gestation in 71 women with subsequent pre-eclampsia and 71 controls matched for age, parity and first trimester body mass index.. Leptin and TGF-beta(1) concentrations were lower and PAI-2 concentration higher in women destined to develop pre-eclampsia relative to controls (leptin: median (25-75 percentiles): 19.0 (14.5-29.0) vs 25.0 (16.0-35.0) ng/ml (p =0.03), TGF-beta(1): 3.2 (2.0-6.1) vs 5.3 (3.8-7.1) ng/ml (P=0.01) and PAI-2: 78.8 (65.1-118.1) vs 67.6 (61.6-79.6) ng/ml (P=0.002)). OR (95 per cent CI) for pre-eclampsia for women in the upper quartile compared to women in the lower quartile were: leptin: 0.2 (0.03-0.7), TGF-beta(1): 0.2 (0.08-0.7) and PAI-2: 3.1 (1.2-8.2).. Altered plasma concentrations levels of factors involved in the process of placentation in women destined to develop pre-eclampsia, indicate that disturbed formation of the uteroplacental unit is reflected in the maternal circulation before 20 weeks' gestation.

Topics: Adult; Biomarkers; Female; Humans; Leptin; Odds Ratio; Placentation; Plasminogen Activator Inhibitor 2; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Second; Prospective Studies; Transforming Growth Factor beta; Transforming Growth Factor beta1

Midtrimester amniotic fluid cytokines may reflect the function of the maternal immune system in the maternal-fetal interface and thus be predictive of pre-eclampsia. We determined the concentrations of interleukin (IL)-6, IL-8, IL-10, IL-11, IL-12, IL-15, tumour necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta in amniotic fluid at 14-16 weeks of gestation from women with normal pregnancies and from those who subsequently developed severe pre-eclampsia. The concentrations of the cytokines in amniotic fluid did not significantly differ between patients and normal controls. The median concentration of IL-6 was 950 pg/ml in normal pregnant women and 578 pg/ml in the patient group. The median concentration of IL-8 was 606 pg/ml in normal controls and 294 pg/ml in the patient group. The levels of IL-6, IL-8 and TGF-beta correlated positively with each other. TNF-alpha concentrations were low and similar in both groups. IL-10 and IL-12 were detected at very low levels in 37 and 7% of the samples, respectively. No difference was found in IL-15 concentrations between the groups. IL-11 was found only at low levels in both groups. Although none of the cytokines measured was predictive of pre-eclampsia, this study provides information of cytokines in amniotic fluid during the period when the spiral arteries are remodelled.

Topics: Adult; Amniotic Fluid; Case-Control Studies; Cytokines; Female; Humans; Inflammation Mediators; Interleukin-12; Interleukin-15; Interleukins; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Second; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Normal human pregnancy depends on physiological transformation of spiral arteries by invasive trophoblasts. Preeclampsia (PE) and fetal growth restriction (FGR) are associated with impaired trophoblast invasion and spiral artery transformation. Recent studies have suggested that transforming growth factor (TGF)-beta3 is overexpressed in the placenta of PE patients and that this may be responsible for failed trophoblast invasion. There are, however, no studies on TGF-betas in the placenta in FGR or in the placental bed in PE or FGR. In this study we have used immunohistochemistry, Western blot analysis, and enzyme-linked immunosorbent assay to examine the expression of TGF-beta1, TGF-beta2, and TGF-beta3 in placenta and placental bed of pregnancies complicated by PE and FGR and matched control pregnancies. The results show that TGF-beta1, -beta2, and -beta3 are not expressed in villous trophoblasts but are present within the placenta. TGF-beta1, -beta2, and, to a much lesser extent, TGF-beta3 were present within the placental bed but only TGF-beta2 was present in extravillous trophoblast. No changes in expression of either isoform were found in placenta or placental bed in PE or FGR compared with normal pregnancy. These data are not consistent with overexpression of TGF-beta3 being responsible for failed trophoblast invasion in PE. Our findings suggest that the TGF-betas do not have a pathophysiological role in either PE or FGR.

Topics: Adult; Biopsy; Blotting, Western; Enzyme-Linked Immunosorbent Assay; Female; Fetal Growth Retardation; Humans; Immunohistochemistry; Placenta; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Third; Reference Values; Transforming Growth Factor beta

Preeclampsia, the major cause of maternal morbidity and mortality in developed countries, is associated with abnormalities of placenta function due to shallow invasion of the maternal decidua by trophoblasts. Data suggest that TGF-beta may play a role in inhibiting trophoblast outgrowth or invasion, or both. We report that placental TGF-beta 3 expression is high in early pregnancy but falls at around 9 weeks' gestation. This pattern is inversely correlated with trophoblast outgrowth and fibronectin synthesis, markers of early trophoblast differentiation toward an invasive phenotype. We demonstrate that TGF-beta 3 is overexpressed in preeclamptic placentae. In contrast to control placentae, explants from preeclamptic pregnancies fail to exhibit spontaneous invasion in vitro. Significantly, antisense-induced inhibition of TGF-beta 3 expression, and inhibition of TGF-beta 3 activity with antibodies, induces the formation of columns of trophoblast cells, which migrate out of the explant into the underlying Matrigel. To our knowledge, this is the first demonstration that the hypoinvasive placental phenotype characteristic of preeclampsia can be essentially normalized in vitro by biochemical manipulation. We speculate that a failure to downregulate expression of TGF-beta 3 at around 9 weeks' gestation results in shallow trophoblast invasion and predisposes the pregnancy to preeclampsia.

Topics: Cell Differentiation; Chorionic Villi; Down-Regulation; Extracellular Space; Female; Humans; Organ Culture Techniques; Phenotype; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, First; Protein Isoforms; Transforming Growth Factor beta; Trophoblasts

In order to investigate the pathophysiological significance of anti-endothelial cell antibody (AECA) in pre-eclampsia, the effects of AECA on endothelin-1 (ET-1) and prostaglandin I2 (PGI2) release from cultured human umbilical vein endothelial cells (HUVEC) was evaluated. Serum samples were taken from 85 pre-eclamptic and 20 normal pregnant women. Anti-endothelial cell antibody was measured by ELISA using HUVEC. The release of ET-1 and 6-keto PGF1-alpha, a stable metabolite of PGI2, from HUVEC were evaluated after incubation with IgG-AECA-positive sera and IgG isolated from AECA-positive sera. The incidence of IgG- and IgM-AECA was 24.7 and 8.2%, respectively. The release of ET-1, in the medium containing IgG-AECA-positive sera was significantly greater than in the medium containing IgG-AECA-negative sera. There was significant correlation between the levels of IgG-AECA and the release of ET-1 from endothelial cells. The ET-1 release by IgG isolated from AECA-positive sera was greater than that from AECA-negative sera. However, the release of 6-keto PGF1-alpha by AECA-positive sera was not significantly different from that of AECA-negative sera. It is concluded that IgG-AECA in pre-eclampsia increases ET-1 release from endothelial cells and that AECA may affect local vascular function in this disorder.

Topics: 6-Ketoprostaglandin F1 alpha; Autoantibodies; Cells, Cultured; Dose-Response Relationship, Immunologic; Endothelin-1; Endothelium, Vascular; Female; Humans; Immunoglobulin G; Interleukin-1; Pre-Eclampsia; Pregnancy; Time Factors; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

This study was performed to investigate the possible association between preeclampsia and the plasma concentrations of Lp(a) lipoprotein and TGF-beta1 in a large series of patients. Additionally, correlation between the concentrations of these molecules and the severity of preeclampsia or fetal growth retardation was evaluated. Following clinical examination and biochemical analyses, both electroimmunoassay and RIA technique were used for quantitative determinations of plasma Lp(a) lipoprotein. ELISA technique was used to measure the active form of TGF-beta1 in plasma of pregnant normotensive and preeclamptic women. We examined 154 women with preeclampsia (preeclampsia group) and 76 healthy, pregnant normotensive women (control group). The preeclampsia group was further divided into the following subgroups: mild preeclampsia, severe preeclampsia and preeclampsia with fetal growth retardation. Plasma levels of Lp(a) lipoprotein were lower in the total preeclampsia group as well as in all preeclampsia subgroups (5.45+/-7.41, 5.58+/-8.02, 5.08+/-5.38, and 4.32+/-5.28 mg/dl in the total preeclampsia group, and in subgroups with mild preeclampsia, severe preeclampsia, and preeclampsia with fetal growth retardation, respectively) than in the control group (7.84+/-9.26 mg/dl) as determined by quantitative electroimmunoassay. Corresponding results were obtained with a radioimmunoassay (166.03+/-200.2 U/l in the total preeclampsia group vs. 229.18+/-257.7 U/l in controls). There was good correlation between the two methods used for Lp(a) lipoprotein measurement. The differences between controls and the total preeclampsia group as well as each preeclampsia subgroup were statistically significant by a non-parametric test (one-way Kruskal-Wallis test). Plasma concentrations of the active form of TGF-beta1 were increased in all preeclampsia subgroups as well as in the total group (5.63+/-1.68 ng/ml) compared to controls (4.67+/-1.33 ng/ml). This increase in TGF-beta1 was statistically highly significant. Plasma concentrations of Lp(a) lipoprotein and the active form of TGF-beta1 did not differ significantly between the preeclampsia subgroups. The outcome of this study may suggest involvement of both parameters in the pathophysiology of preeclampsia and may substantiate the notion of a multifactorial etiology of the disease.

Topics: Adult; Arteriosclerosis; Birth Weight; Blood Pressure; Female; Fetal Growth Retardation; Gestational Age; Humans; Hypertension; Lipoprotein(a); Maternal Age; Pre-Eclampsia; Pregnancy; Risk Factors; Transforming Growth Factor beta