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1.
Gene ; 749: 144721, 2020 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-32360842

RESUMO

Fetal development is critically dependent on the efficiency of the placenta. Porcine trophoblast cell proliferation and invasion have crucial roles in placental fold development, which is one of the essential events determining placental efficiency. The membrane serine proteinase inhibitor hepatocyte growth factor activator inhibitor-1 (HAI-1) can regulate cellular invasion and motility in different types of epithelial cells, including trophoblast cells in mice. This work used quantitative polymerase chain reaction (qPCR) and immunohistochemistry to compare the expression level and location of HAI-1 in the placenta on gestational days 26, 50, and 95 in Yorkshire and Meishan pigs. The role of HAI-1 in porcine trophoblast cell (PTr2) proliferation, invasion, and migration in vitro was investigated by analyzing the effects of HAI-1 gene silencing or overexpression. Polymorphism in the HAI-1gene was detected to determine associations between the genotype and piglet birth weight in 400 healthy pure-bred Yorkshire piglets. qPCR results showed that HAI-1 mRNA levels significantly increased (P < 0.01) between gestational days 26 and 50 and then decreased (P < 0.01) between days 50 and 95 in both Meishan and Yorkshire pigs. Immunohistochemical analysis showed that HAI-1 protein was strongly expressed by the high columnar trophoblast cells located at the top of the placental folds with low proliferative and invasion capacities. However, it was expressed at very low levels in cuboidal trophoblast cells located at the side and base of the placental folds with high proliferative and invasion capacities. In vitro experiments indicated that HAI-1 had the ability to reduce the proliferation, invasion and migration of trophoblast cells. In addition, one single-nucleotide polymorphism (SNP) of HAI-1 showed a significant association (P < 0.05) with piglet birth weight. These results revealed that HAI-1 could be a vital molecule in placental folds development by regulating trophoblast proliferation and invasion in pigs.


Assuntos
Placenta/metabolismo , Placentação , Proteínas Secretadas Inibidoras de Proteinases/fisiologia , Sus scrofa/embriologia , Trofoblastos/fisiologia , Animais , Linhagem Celular , Movimento Celular , Proliferação de Células , Feminino , Placentação/genética , Polimorfismo de Nucleotídeo Único , Gravidez , Proteínas Secretadas Inibidoras de Proteinases/genética , Proteínas Secretadas Inibidoras de Proteinases/metabolismo , RNA Mensageiro/metabolismo , Sus scrofa/genética , Sus scrofa/metabolismo , Trofoblastos/citologia
2.
Environ Pollut ; 264: 114790, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32417587

RESUMO

The ongoing transition to renewable fuel sources has led to increased use of wood and other biomass fuels. The physiochemical characteristics of biomass combustion derived aerosols depends on appliances, fuel and operation procedures, and particles generated during incomplete combustion are linked to toxicity. Frequent indoor wood burning is related to severe health problems such as negative effects on airways and inflammation, as well as chronic hypoxia and pathological changes in placentas, adverse pregnancy outcome, preterm delivery and increased risk of preeclampsia. The presence of combustion-derived black carbon particles at both the maternal and fetal side of placentas suggests that particles can reach the fetus. Air pollution particles have also been shown to inhibit trophoblast migration and invasion, which are vital functions for the development of the placenta during the first trimester. In this study we exposed a placental first trimester trophoblast cell line to wood smoke particles emitted under Nominal Burn rate (NB) or High Burn rate (HB). The particles were visible inside exposed cells and localized to the mitochondria, causing ultrastructural changes in mitochondria and endoplasmic reticulum. Exposed cells showed decreased secretion of the pregnancy marker human chorionic gonadotropin, increased secretion of IL-6, disrupted membrane integrity, disrupted proliferation and contained specific polycyclic aromatic hydrocarbons (PAHs) from the particles. Taken together, these results suggest that wood smoke particles can enter trophoblasts and have detrimental effects early in pregnancy by disrupting critical trophoblast functions needed for normal placenta development and function. This could contribute to the underlying mechanisms leading to pregnancy complications such as miscarriage, premature birth, preeclampsia and/or fetal growth restriction. This study support the general recommendation that more efficient combustion technologies and burning practices should be adopted to reduce some of the toxicity generated during wood burning.


Assuntos
Fumaça , Trofoblastos , Proliferação de Células , Feminino , Humanos , Recém-Nascido , Inflamação , Gravidez , Madeira
3.
Nan Fang Yi Ke Da Xue Xue Bao ; 40(1): 79-86, 2020 Jan 30.
Artigo em Chinês | MEDLINE | ID: mdl-32376568

RESUMO

OBJECTIVE: To investigate the roles of microRNA (miR)-34a-5p and cyclin-dependent kinase (CDK) 6 in the regulation of cell viability, apoptosis and invasion of human placental trophoblastic cells and the relationship between miR-34a-5p and CDK6. METHODS: We examined the expression of miR-34a-5p using RT-qPCR in cultured human trophoblast HTR-8/Svneo cells and human choriocarcinoma cell lines BeWo and JEG-3HTR-8/Svneo. HTR-8/Svneo cells transfected with a miR-34a-5p-mimic, the miR-34a-5p-inhibitor, or pcDNA-CDK6 along with the mimic group were analyzed for changes in cell proliferation using MTT assay; the apoptosis of the cells were assessed by detecting caspase 3 activity and cleaved caspase 3 protein expression, and the cell invasion was evaluated using Transwell assay. Western blotting was used to determine the protein levels of CDK6, cleaved caspase 3, and MMP-9 in the cells. The interaction between CDK6 and miR-34a-5p analyzed using a luciferase reporter assay. RESULTS: Transfection with the miR-34a-5p mimic significantly reduced the viability (P=0.000), suppressed the invasion (P=0.049), enhanced the cell apoptosis (P=0.018), down-regulated the expressions of MMP-9 (P=0.004) and CDK6 (P=0.014), and up-regulated caspase 3 activity (P=0.018) and cleaved caspase 3 expression (P=0.003) in cultured HTR-8/Svneo cells. CDK6 was confirmed as one of the target gene of miR-34a-5p. Transfection with pcDNA-CDK6 significantly reversed the effects of miR- 34a-5p overexpression on the cell viability (P=0.000), apoptosis (P=0.015), and invasion (P=0.046). Treatment of the cells with insulin-like growth factor 1 (IGF-1), an activator of the PI3K/AKT pathway, also significantly attenuated the effects of miR-34a- 5p overexpression on the cell viability (P=0.011), apoptosis (P=0.004), and invasion (P=0.002). CONCLUSIONS: miR-34a-5p promotes apoptosis and inhibits the viability and invasion of human placental trophoblastic cells by down-regulating CDK6 and inactivating the PI3K/AKT pathway.


Assuntos
Apoptose , MicroRNAs/genética , Transdução de Sinais , Trofoblastos/citologia , Proliferação de Células , Células Cultivadas , Quinase 6 Dependente de Ciclina/metabolismo , Feminino , Humanos , Fosfatidilinositol 3-Quinases/metabolismo , Gravidez , Proteínas Proto-Oncogênicas c-akt/metabolismo
4.
Life Sci ; 254: 117781, 2020 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-32407842

RESUMO

AIMS: Previous evidence has demonstrated that oxidative stress is related to the pathogenesis of missed abortion (MA), but the specific mechanism remains obscure. The adaptor protein APPL1 is one of the differential proteins in chorionic trophoblasts. Thus, this study aimed to assess the potential influence of APPL1 on oxidative stress responses as well the possible molecular mechanisms involving in MA. MAIN METHODS: In the present study, the chorionic trophoblasts and the HTR-8/SVneo cell line were researched in vitro. Small interfering RNA (siRNA) was used to suppress the expression of APPL1. The fluorescent probes DHE and DCFH-DA were used to assess the intracellular reactive oxidative species (ROS). The activity of superoxide dismutase (SOD) was determined. Apoptosis was detected by TUNEL and flow cytometry. Cell viability was determined using Cell Counting Kit-8. Protein expression was detected by immunohistochemistry, western blotting, and reverse transcription-quantitative PCR. KEY FINDINGS: The application of oxidant in normal chorionic trophoblasts induced cell death and overproduction of ROS, which was consistent with MA. In addition, knockdown of APPL1 in HTR-8/SVneo cells resulted in increased ROS and apoptosis, which could be rescued by pretreatment with antioxidants. Mechanistically, we report that overproduction of ROS in trophoblasts and disturbed SOD, APPL1 and Nrf2/HO-1 antioxidant responses constitute important contributors to apoptosis. SIGNIFICANCE: Our results suggest that APPL1 has antioxidant properties that suppress oxidative stress and apoptosis via the Nrf2/HO-1 pathway. Moreover, antioxidant N-acetylcysteine (NAC) effectively restored the impaired antioxidative defense system elicited by excess ROS, as a potential therapeutic reagent for MA.


Assuntos
Aborto Retido/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Apoptose/fisiologia , Heme Oxigenase-1/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/fisiologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Feminino , Técnicas de Silenciamento de Genes , Humanos , Gravidez , RNA Interferente Pequeno/farmacologia , Superóxido Dismutase/metabolismo , Trofoblastos/metabolismo
5.
Cell Prolif ; 53(5): e12802, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32291850

RESUMO

OBJECTIVES: During human pregnancy, the endothelial cells of the uterine spiral arteries (SPA) are extensively replaced by a subtype of placental trophoblasts, endovascular extravillous trophoblasts (enEVTs), thus establishing a placental-maternal circulation. On this pathway, foetus-derived placental villi and enEVTs bath into the maternal blood that perfuses along SPA being not attacked by the maternal lymphocytes. We aimed to reveal the underlying mechanism of such immune tolerance. METHODS: In situ hybridization, immunofluorescence, ELISA and FCM assay were performed to examine TGF-ß1 expression and distribution of regulatory T cells (Tregs) along the placental-maternal circulation route. The primary enEVTs, interstitial extravillous trophoblasts (iEVTs) and decidual endothelial cells (dECs) were purified by FACS, and their conditioned media were collected to treat naïve CD4+ T cells. Treg differentiation was measured by FLOW and CFSE assays. RESULTS: We found that enEVTs but not iEVTs or dECs actively produced TGF-ß1. The primary enEVTs significantly promoted naïve CD4+ T-cell differentiation into immunosuppressive FOXP3+ Tregs, and this effect was dependent on TGF-ß1. In recurrent spontaneous abortion (RSA) patients, an evidently reduced proportion of TGF-ß1-producing enEVTs and their ability to educate Tregs differentiation were observed. CONCLUSIONS: Our findings demonstrate a unique immune-regulatory characteristic of placental enEVTs to develop immune tolerance along the placental-maternal circulation. New insights into the pathogenesis of RSA are also suggested.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Diferenciação Celular/imunologia , Placenta/imunologia , Linfócitos T Reguladores/imunologia , Trofoblastos/imunologia , Adulto , Animais , Células Endoteliais/imunologia , Feminino , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Gravidez , Fator de Crescimento Transformador beta1/imunologia
6.
Life Sci ; 253: 117668, 2020 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-32320706

RESUMO

AIMS: Preeclampsia (PE) accounts for the foremost cause of maternal and fetal mortality worldwide, whereas, there are no effective treatments for the disease yet. Long non-coding RNAs (lncRNAs) play critical roles in various human disorders, including PE. Here, we identified an up-regulated lncRNA HOTAIR, and explored its underlying mechanisms in PE. MAIN METHODS: qRT-PCR analysis was used to examine HOTAIR expression in PE tissues and cell lines. Trophoblast proliferation was examined by colony formation and 5-Ethynyl-2'-deoxyuridine (EdU) incorporation assays. Trophoblast migration and invasion was determined by transwell and wound healing assays. Bioinformatics analysis was performed to verify the regulation HOTAIR on miRNAs. The interaction between HOTAIR and EZH2 was detected using RNA immunoprecipitation assay (RIP). Chromatin immunoprecipitation (CHIP) assay was also performed to verify that the negative regulation of HOTAIR on miR-106a was dependent on the epigenetic repressor EZH2. KEY FINDINGS: HOTAIR was up-regulated in PE placenta tissues, which repressed the proliferation, migration and invasion of trophoblast cells. HOTAIR significantly repressed miR-106a expression and the reduced miR-106a level was also observed in placentas from PE patients. Additionally, miR-106a mimic enhanced the migration and invasion of trophoblast cells. Further mechanistic analyses implied that the action of HOTAIR is moderately attributable to its repression of miR-106a via association with EZH2. SIGNIFICANCE: High level of HOTAIR repressed the proliferation, migration and invasion of trophoblast cells through targeting miR-106 in an EZH2-dependent manner, which may provide new insights into the roles of HOTAIR and miR-106a as potential regulators in PE.


Assuntos
Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , MicroRNAs/metabolismo , Pré-Eclâmpsia/metabolismo , RNA Longo não Codificante/metabolismo , Sequência de Bases , Linhagem Celular , Proliferação de Células , Progressão da Doença , Repressão Epigenética , Feminino , Humanos , Placenta/metabolismo , Gravidez , Trofoblastos/citologia , Regulação para Cima
7.
Chemosphere ; 254: 126785, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32334250

RESUMO

Fetal overexposure to active glucocorticoid (GC) is the major cause for fetal growth restriction (FGR). This study investigated the influences of cadmium (Cd) exposure on active GC and its mechanism in placental trophoblasts. Pregnant mice were exposed to CdCl2 (4.5 mg/kg, i.p.). Human JEG-3 cells were treated with CdCl2 (0-20 µM). Prenatal Cd exposure significantly increased active GC level in amniotic fluid and placenta. Similarly, Cd treatment also elevated active GC level in medium. Expectedly, the expression of 11ß-HSD2 protein was markedly downregulated in Cd-exposed placental trophoblasts. We further found that Cd activated the PERK/p-eIF2α signaling pathway in placental trophoblasts. Mechanistically, PERK siRNA pretreatment completely blocked PERK/p-eIF2α signaling, and thereby restoring Cd-downregulated 11ß-HSD2 protein expression in human placental trophoblasts. We further found that N-acetylcysteine, a well-known antioxidant, obviously reversed Cd-downregulated 11ß-HSD2 protein expression by inhibiting p-PERK/p-eIF2α signaling in placental trophoblasts. Overall, our data suggest that Cd activates the PERK/p-eIF2α signaling, down-regulates the protein expression of 11ß-HSD2, and thereby elevating active GC level in placental trophoblast.


Assuntos
11-beta-Hidroxiesteroide Desidrogenase Tipo 2/metabolismo , Cádmio/toxicidade , Poluentes Ambientais/toxicidade , Animais , Cádmio/metabolismo , Linhagem Celular Tumoral , Regulação para Baixo , Fator de Iniciação 2 em Eucariotos/metabolismo , Feminino , Retardo do Crescimento Fetal , Glucocorticoides/metabolismo , Humanos , Camundongos , Placenta/metabolismo , Gravidez , Trofoblastos/fisiologia
8.
Life Sci ; 251: 117625, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32247003

RESUMO

OBJECTIVE: The present study was designed to investigate whether the novel peptide cysteine-based peptide (Cys-peptide) had protective effects on preeclamptic animal and cell models. METHODS: We investigated effects of Cys-peptide on (1) preeclamptic symptoms (e.g. hypertension, proteinuria, fetal growth restriction (FGR)) in preeclampia-like rat models induced by lipopolysaccharides (LPS), (2) TNFα-induced cytotoxicity of human umbilical vascular endothelial cells (HUVECs) and HTR-8 cells (an immortalised human trophoblast cell line), (3) endothelial dysfunction and injured angiogenesis, (4) migration and invasion of trophoblast cells induced by TNFα. RESULTS: Cys-peptide ameliorated LPS-induced hypertension, proteinuria and FGR and other PE symptoms in preeclampia-like rat models. In addition, Cys-peptide attenuated TNFα-induced cytotoxicity by decreasing soluble fms-like tyrosine kinase-1 (sFlt-1), endothelin-1 (ET-1) and tissue plasminogen activator (tPA) mRNA expression in both cells. Furthermore, Cys-peptide restored endothelial dysfunction and rescued angiogenesis caused by TNFα in vitro. Importantly, Cys-peptide could reverse insufficient ability to invade and migrate of trophoblast cells. CONCLUSIONS: These results suggest Cys-peptide can play beneficial roles in preeclampsia-like rat and cell models. Therefore, we propose that Cys-peptide is probably a novel therapeutic candidate for PE.


Assuntos
Cisteína/química , Retardo do Crescimento Fetal/prevenção & controle , Peptídeos/administração & dosagem , Pré-Eclâmpsia/prevenção & controle , Animais , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Modelos Animais de Doenças , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Hipertensão/prevenção & controle , Peptídeos/química , Peptídeos/farmacologia , Pré-Eclâmpsia/fisiopatologia , Gravidez , Proteinúria/prevenção & controle , Ratos , Ratos Sprague-Dawley , Trofoblastos/metabolismo , Fator de Necrose Tumoral alfa/administração & dosagem
9.
Clin Sci (Lond) ; 134(6): 593-607, 2020 03 27.
Artigo em Inglês | MEDLINE | ID: mdl-32129439

RESUMO

Small extracellular vesicles (sEVs) released from the extravillous trophoblast (EVT) are known to regulate uterine spiral artery remodeling during early pregnancy. The bioactivity and release of these sEVs differ under differing oxygen tensions and in aberrant pregnancy conditions. Whether the placental cell-derived sEVs released from the hypoxic placenta contribute to the pathophysiology of preeclampsia is not known. We hypothesize that, in response to low oxygen tension, the EVT packages a specific set of proteins in sEVs and that these released sEVs interact with endothelial cells to induce inflammation and increase maternal systemic blood pressure. Using a quantitative MS/MS approach, we identified 507 differentially abundant proteins within sEVs isolated from HTR-8/SVneo cells (a commonly used EVT model) cultured at 1% (hypoxia) compared with 8% (normoxia) oxygen. Among these differentially abundant proteins, 206 were up-regulated and 301 were down-regulated (P < 0.05), and they were mainly implicated in inflammation-related pathways. In vitro incubation of hypoxic sEVs with endothelial cells, significantly increased (P < 0.05) the release of GM-CSF, IL-6, IL-8, and VEGF, when compared with control (i.e. cells without sEVs) and normoxic sEVs. In vivo injection of hypoxic sEVs into pregnant rats significantly increased (P < 0.05) mean arterial pressure with increases in systolic and diastolic blood pressures. We propose that oxygen tension regulates the release and bioactivity of sEVs from EVT and that these sEVs regulate inflammation and maternal systemic blood pressure. This novel oxygen-responsive, sEVs signaling pathway, therefore, may contribute to the physiopathology of preeclampsia.


Assuntos
Citocinas/metabolismo , Vesículas Extracelulares/química , Hipóxia/fisiopatologia , Oxigênio/metabolismo , Pré-Eclâmpsia/fisiopatologia , Animais , Pressão Arterial , Pressão Sanguínea , Citocinas/genética , Células Endoteliais/química , Células Endoteliais/metabolismo , Vesículas Extracelulares/metabolismo , Feminino , Humanos , Hipóxia/genética , Hipóxia/metabolismo , Oxigênio/análise , Pré-Eclâmpsia/genética , Pré-Eclâmpsia/metabolismo , Gravidez , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas em Tandem , Trofoblastos/química , Trofoblastos/metabolismo
10.
PLoS One ; 15(3): e0229949, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32182245

RESUMO

We present a two-tiered microchip system to capture and retrieve rare cells from blood samples with high purity. The first module of the system is a high throughput microfluidic interface that is used to immunomagnetically isolate targeted rare cells from whole blood, and discard > 99.999% of the unwanted leukocytes. The second module is a microwell array that furthers the purification by magnetically guiding each cell into a separate well concurrently, and allows individual retrieval of each cell. We demonstrate the design of the system as well as its characterization by experiments using model cell lines that represent circulating fetal trophoblasts. Our results show that single cells can be retrieved with efficiencies and purities as high as 100% within 145 mins.


Assuntos
Separação Celular , Procedimentos Analíticos em Microchip , Neoplasias/sangue , Análise de Célula Única , Linhagem Celular Tumoral , Humanos , Leucócitos/citologia , Análise em Microsséries , Microfluídica/métodos , Neoplasias/patologia , Trofoblastos/citologia
11.
Bratisl Lek Listy ; 121(3): 225-229, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32115981

RESUMO

AIM: Nicotine at high concentrations induces apoptosis in trophoblastic cells through induction of cell cytotoxicity and Reactive Oxygen Species (ROS). Methamphetamine in low dose has pharmaceutical properties. It seems that this components in low dose can protect the trophoblastic cells from nicotine-induced cell death. METHOD: Trophoblastic (JEG-3) cells grown in DMEM culture medium. MTT assay test detected the cell viability and Lactate Dehydrogenase test measured the cells cytotoxicity. Griess reaction was used for NO production analysis. Cell migration traced by wounding technique. Human Cytokine Array Focused 13-plex was also used for analysis of IL-1α, IL-1ß, IL-6, INFγ, and TNFα pre-inflammatory cytokines. RESULTS: Methamphetamine, in very low dose (pM), increased the cell viability and NO production, and decreased cell cytotoxicity, IL-1α, IL-1ß, IL-6, INFγ, and TNFα pre-inflammatory cytokines of JEG-3 cell which were exposed to high dose of nicotine, respectively. Cell migration was enhanced by low dose of methamphetamine in JEG-3 cells. CONCLUSION: Methamphetamine in very low dose suppressed the JEG-3 cell death induced by high dose of nicotine (Fig. 5, Ref. 48) Keywords: methamphetamine, nicotine, cell death, NO.


Assuntos
Dopaminérgicos , Inflamação , Metanfetamina , Trofoblastos , Linhagem Celular Tumoral , Sobrevivência Celular , Dopaminérgicos/farmacologia , Humanos , Inflamação/tratamento farmacológico , Metanfetamina/farmacologia , Nicotina/toxicidade , Trofoblastos/efeitos dos fármacos
12.
Gene ; 741: 144533, 2020 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-32145327

RESUMO

BACKGROUND: Syncytin-1 and syncytin-2 which are endogenous retroviral genes products play a great role in syncytialization during trophoblast differentiation in normal placental tissues. In aneuploidic placentas due to the low level of pregnancy-induced hormones an alteration was occurred in the syncytialization process, while in the presence of cytogenetically abnormal karyotype the effect of syncytin gene expression levels on syncytialization process and in occured to spontaneous abortions is not clear. To reveal this, we investigated in syncytin-1 and syncytin-2 genes expression levels of chromosomally abnormal and normal trophophoblastic tissues and we also discussed the effect of the syncytin gene expression levels to the occurense of the spontaneous abortion. MATERIAL AND METHODS: To each one of the trophoblastic cells; cultivation, harvesting, banding, and analysis were performed and the chromosomes were classified according to the presence of abnormality and normal XY constitution. To exclude the maternal decidual cell contamination, female karyotyped abortion materials were omitted in control group. The patient group consisted of thirty six placental tissues including trisomy 16 (n = 10), triploidy (n = 9), monosomy X (n = 9), trisomy 21 (n = 5) and trisomy 7 (n = 3). The control group was consisting twenty placental tissues with XY karyotypes. The some part of the dissected frozen trophoblastic cells were used for RNA isolation and were proceeded to the determination of the expression levels of syncytin-1 and syncytin-2 genes by single-step Real Time PCR. The cDNAs were obtained by probes used in the same PCR stages. The sequence analysis of the syncytin-1 and syncytin-2 genes were performed, and read by the usage of the FinchTV 1.4.0 program. RESULTS: Between the expression levels of syncytin-1 and syncytin-2 genes were statistically difference in the patients and controls. There was a difference (p < 0.0001) between trisomy 7 and other patient groups and controls, regarding to the expression of syncytin-1 gene. Numerous mutations in the syncytin-1 and syncytin-2 genes (on the expression sites) were detected, and the mutation rate was higher in the syncytin-1 gene compared to the syncytin-2 gene in the patient and in the control groups (p < 0.001). CONCLUSION: The results of the study indicate that the expression of the syncytin-2 genes could be altered in the presence of chromosomally abnormal trophoblastic tissues, and these could lead to the loss of pregnancy due to the insufficient syncytialization. In sum, the current research has value for the further studies covering the mechanisms of trophoblast cell fusion, and syncytiotrophoblast regeneration, and thus the pathophysiology of human placental development in the presence of genomic anomaly.


Assuntos
Cariótipo Anormal , Aborto Espontâneo/genética , Produtos do Gene env/economia , Proteínas da Gravidez/economia , Proteínas da Gravidez/genética , Feto Abortado/patologia , Aborto Induzido/métodos , Aborto Espontâneo/patologia , Adulto , Diferenciação Celular/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Cariótipo , Placenta/metabolismo , Gravidez , Trissomia/genética , Trissomia/patologia , Trofoblastos/patologia , Síndrome de Turner/genética , Síndrome de Turner/patologia , Adulto Jovem
13.
Am J Pathol ; 190(5): 970-976, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32084366

RESUMO

Preeclampsia (PE) is a hypertensive disease of pregnancy associated with substantial maternal and fetal morbidity and mortality. CORIN is a transmembrane type II serine protease expressed in cardiomyocytes that converts pro-atrial natriuretic peptide into atrial natriuretic peptide, a cardiac hormone that regulates blood pressure. High levels of soluble CORIN have been reported in PE and are supposed to be cardiac in origin. We hypothesized that during pregnancy soluble CORIN is released by the syncytiotrophoblast and that increased levels of soluble CORIN in preeclampsia originate from placenta. A total of 375 patients (181 PE patients and 194 controls) were analyzed. High levels of soluble CORIN were confirmed in maternal blood from preeclamptic pregnancies compared with controls. Differentiated primary villous cytotrophoblasts showed that CORIN was expressed (mRNA and protein levels) and secreted by trophoblastic cells, mostly by the syncytiotrophoblast. Finally, placental explants showed a significant increase in CORIN production and secretion in PE cases compared with controls. This study showed that CORIN is secreted by trophoblastic cells and that high levels of soluble CORIN in preeclampsia have a placental origin.


Assuntos
Placenta/metabolismo , Pré-Eclâmpsia/metabolismo , Serina Endopeptidases/biossíntese , Feminino , Humanos , Gravidez , Trofoblastos/metabolismo
14.
Clin Sci (Lond) ; 134(5): 459-472, 2020 03 13.
Artigo em Inglês | MEDLINE | ID: mdl-32068238

RESUMO

Antiphospholipid autoantibodies (aPLs), a major maternal risk factor for preeclampsia, are taken into the syncytiotrophoblast where they bind intracellular vesicles and mitochondria. Subsequently, large quantities of extracellular vesicles (EVs) extruded from syncytiotrophoblast into the maternal circulation are altered such that they cause maternal endothelial cell activation. However, the mechanism driving this change is unknown. First trimester placental explants were treated with aPL for 18 h. The EVs were then collected by different centrifugation. The levels of HSP 70, misfolded proteins, caspase 8 activity, and Mixed Lineage Kinase domain-Like (MLKL) were measured in placental explants and EVs. In addition, the levels of TNF-α and CD95 in conditioned medium were also measured. Treating placental explants with aPL caused an increase in levels of HSP 70, misfolded proteins and MLKL in placental explants and EVs. Increased activity of caspase 8 was also seen in placental explants. Higher levels of TNF-α were seen conditioned medium from aPL-treated placental explant cultures. aPLs appear to induce endoplasmic reticulum stress in the syncytiotrophoblast in a manner that involved caspase 8 and TNF-α. To avoid accumulation of the associated misfolded proteins and MLKL, the syncytiotrophoblast exports these potentially dangerous proteins in EVs. It is likely that the dangerous proteins that are loaded into placental EVs in preeclampsia contribute to dysfunction of the maternal cells.


Assuntos
Anticorpos Antifosfolipídeos/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Vesículas Extracelulares/metabolismo , Placenta/efeitos dos fármacos , Trofoblastos/efeitos dos fármacos , Caspase 8/metabolismo , Feminino , Proteínas de Choque Térmico HSP70/metabolismo , Humanos , Placenta/metabolismo , Pré-Eclâmpsia/metabolismo , Gravidez , Primeiro Trimestre da Gravidez , Proteínas Quinases/metabolismo , Técnicas de Cultura de Tecidos , Trofoblastos/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
15.
Virchows Arch ; 477(1): 73-81, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32025822

RESUMO

Villitis of unknown etiology (VUE) and chronic deciduitis with plasma cells (CD) are supposed to be non infectious placental lesions caused by a pathologic immune reaction similar to a host versus graft mechanism. In some investigations, infection of human trophoblastic cells with human papilloma virus (HPV) has been described, and a relationship with miscarriage, preeclampsia, and chronic inflammatory placental lesions has been suspected. Infection with enterovirus, especially Coxsackievirus, has been observed in cases with spontaneous abortion and adverse perinatal outcome, respectively. We investigated 20 cases with VUE and 30 cases with chronic deciduitis with plasma cells. The placenta specimens were analyzed for expression of HPV capsid protein by immunohistochemistry, for presence of HPV DNA via polymerase chain reaction (PCR), and for presence of enterovirus mRNA using RT-PCR, respectively. VUE was associated with maternal diseases: atopic lesions in 21%, other autoimmune diseases in 15.5%, and obesity in 31.5%, respectively. Birth weight below the 10th percentile was detected in 63% of the cases with VUE. Chronic deciduitis was associated with preterm labor and preterm premature rupture of membranes (26%). Intrauterine fetal demise occurred in 5 cases with CD (18.5%). HPV DNA, HPV capsid protein, and enterovirus mRNA were not detected in all investigated VUE or CD cases. Our investigations show that a causal role for enterovirus and human papilloma virus in the development of VUE and CD is unlikely. Therefore, HPV vaccination is unlikely to reduce the incidence of VUE and CD in the future.


Assuntos
Corioamnionite/etiologia , Vilosidades Coriônicas/patologia , Papillomaviridae/patogenicidade , Placenta/virologia , Adulto , Corioamnionite/patologia , Corioamnionite/virologia , Infecções por Enterovirus/etiologia , Feminino , Humanos , Recém-Nascido , Placenta/patologia , Doenças Placentárias/etiologia , Doenças Placentárias/patologia , Gravidez , Trofoblastos/patologia , Trofoblastos/virologia
16.
Sheng Li Xue Bao ; 72(1): 91-104, 2020 Feb 25.
Artigo em Chinês | MEDLINE | ID: mdl-32099987

RESUMO

The onset of tight connection between embryo and uterine endometrium terms "embryo implantation", the beginning and a key step of mammalian pregnancy. Defective implantation leads to failure of pregnancy and infertility. In recent years, along with the technological advance, researches on embryo implantation have achieved great advances. This paper reviews the key research achievements that have been reached in the last decade in the field of embryo implantation, focusing on the changes, roles, and underlying mechanisms of both luminal and glandular epithelia during implantation process, as well as their interactions with embryo trophoblast cells and endometrial stromal cells.


Assuntos
Implantação do Embrião , Endométrio/fisiologia , Animais , Feminino , Gravidez , Células Estromais/fisiologia , Trofoblastos/fisiologia , Útero/fisiologia
17.
Sheng Li Xue Bao ; 72(1): 115-124, 2020 Feb 25.
Artigo em Chinês | MEDLINE | ID: mdl-32099989

RESUMO

Placenta serves as a temporary fetal organ, which mediates maternal-fetal crosstalk and intrauterine fetal growth. Placental defensive barrier is a fundamental physiological function, which balances maternal immune tolerance to the fetus and resistance to pathogens. This review summarizes the latest research progress on the mechanisms of placental barrier formation from the view of placental development. Recent discoveries have shed light on the cellular and molecular properties of placental defensive mechanisms in syncytiotrophoblast, including autophagy, exosome mediated anti-pathogenic pathways, cell-cell junctions and cytoskeleton networks. We also present an overview of placental barrier dysfunction and its implications in intrauterine TORCH infections.


Assuntos
Troca Materno-Fetal , Placenta/fisiologia , Trofoblastos/fisiologia , Autofagia , Citoesqueleto , Exossomos , Feminino , Desenvolvimento Fetal , Feto , Humanos , Gravidez
18.
Arkh Patol ; 82(1): 23-29, 2020.
Artigo em Russo | MEDLINE | ID: mdl-32096487

RESUMO

OBJECTIVE: To identify the structural and immunohistochemical features of the placentas and the placental sites after in vitro fertilization (IVF) with a donor egg (surrogate motherhood). SUBJECT AND METHODS: Morphological and immunohistochemical studies were performed on the placental (a placental disk) and placental bed materials obtained after caesarean delivery. The investigation enrolled 26 patients whose pregnancy occurred with IVF with a donor egg according to the surrogacy (IVF-S) program. A comparison group included 13 patients whose pregnancy occurred after IVF with their own eggs. An immunohistochemical study was conducted on paraffin sections made from biopsy material; mouse antibodies to total cytokeratin (clone AE1/AE3, 'Dako'), HLA-DR (clone TAL.1B5, 'Dako'), and CD138 (clone MI15, 'Dako') were used as primary antibodies. RESULTS: The histological examination of the placentas in the IVF-S group showed the high incidence of central ischemic heart attacks (69%), dissociated cotyledon development (61%), pathological villous immaturity mainly with the predominance of intermediate differentiated villi (46%), and massive perivillous fibrinoid deposition (73%). The obtained differences between with the study and comparison groups were significant (p<0.05). The IVF-S group was characterized by the development of lymphoplasmacytic deciduitis (1.23±0.4 and 0.5±0.3 scores). Examination of the placental site biopsy material in the IVF-S group revealed the following changes: remodeling of the spiral arteries was incomplete in more than 40% of cases, and in 30% of the spiral arteries had no gestational changes. In the comparison group, more than 90% of the spiral arteries were characterized by complete remodeling during pregnancy. There was also an increase in the count of multinucleated trophoblastic giant cells (104.56±4.21 and 65.67±14.45) and HLA-DR positive cells (41.86±5.32 and 29.00±1.87). CONCLUSION: The placentas and the placental sites of the women whose pregnancy occurred with IVF-S are characterized by the development of high lyoplasmacytic deciduitis activity and pronounced placental immune alterations manifested by the high incidence of immune responses at the sites of the closest contact between maternal and fetal tissues. The placental bed exhibited defective spiral artery remodeling, development of chronic inflammatory lesions in the perivascular areas, and an increase in the counts of HLA-DR positive cells and multinucleated trophoblastic giant cells.


Assuntos
Fertilização In Vitro , Placenta , Animais , Córion , Feminino , Humanos , Camundongos , Gravidez , Doadores de Tecidos , Trofoblastos
19.
Proc Natl Acad Sci U S A ; 117(9): 4642-4652, 2020 03 03.
Artigo em Inglês | MEDLINE | ID: mdl-32071231

RESUMO

Placental trophoblast cells are potentially at risk from circulating endocrine-disrupting chemicals, such as bisphenol A (BPA). To understand how BPA and the reputedly more inert bisphenol S (BPS) affect the placenta, C57BL6J mouse dams were fed 200 µg/kg body weight BPA or BPS daily for 2 wk and then bred. They continued to receive these chemicals until embryonic day 12.5, whereupon placental samples were collected and compared with unexposed controls. BPA and BPS altered the expression of an identical set of 13 genes. Both exposures led to a decrease in the area occupied by spongiotrophoblast relative to trophoblast giant cells (GCs) within the junctional zone, markedly reduced placental serotonin (5-HT) concentrations, and lowered 5-HT GC immunoreactivity. Concentrations of dopamine and 5-hydroxyindoleacetic acid, the main metabolite of serotonin, were increased. GC dopamine immunoreactivity was increased in BPA- and BPS-exposed placentas. A strong positive correlation between 5-HT+ GCs and reductions in spongiotrophoblast to GC area suggests that this neurotransmitter is essential for maintaining cells within the junctional zone. In contrast, a negative correlation existed between dopamine+ GCs and reductions in spongiotrophoblast to GC area ratio. These outcomes lead to the following conclusions. First, BPS exposure causes almost identical placental effects as BPA. Second, a major target of BPA/BPS is either spongiotrophoblast or GCs within the junctional zone. Third, imbalances in neurotransmitter-positive GCs and an observed decrease in docosahexaenoic acid and estradiol, also occurring in response to BPA/BPS exposure, likely affect the placental-brain axis of the developing mouse fetus.


Assuntos
Compostos Benzidrílicos/toxicidade , Encéfalo/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Fenóis/toxicidade , Sulfonas/toxicidade , Trofoblastos/efeitos dos fármacos , Animais , Dopamina/metabolismo , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Serotonina/metabolismo , Trofoblastos/metabolismo
20.
Environ Pollut ; 260: 113984, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32041019

RESUMO

1-nitropyrene (1-NP) is a key component of diesel exhaust-sourced fine particulate matter (PM2.5). Our recent study demonstrated that gestational 1-NP exposure caused placental proliferation inhibition and fetal intrauterine growth restriction (IUGR). This study aimed to investigate the role of genotoxic stress on 1-NP-induced placental proliferation inhibition and fetal IUGR. Human trophoblasts were exposed to 1-NP (10 µM). Growth index was reduced and PCNA was downregulated in 1-NP-exposed placental trophoblasts. More than 90% of 1-NP-exposed trophoblasts were arrested in either G0/G1 or G2/M phases. CDK1 and cyclin B, two G2/M cycle-related proteins, and CDK2, a G0/G1 cycle-related protein, were reduced in 1-NP-exposed trophoblasts. Phosphorylated Rb, a downstream molecule of CDK2, was inhibited in 1-NP-exposed trophoblasts. Moreover, DNA double-strand break was observed and γ-H2AX, another indicator of DNA double-strand break, was upregulated in 1-NP-exposed trophoblasts. Phosphorylated ATM, a key molecule of genotoxic stress, and its downstream molecule Chk2 were elevated. By contrast, Cdc25A, a downstream target of Chk2, was reduced in 1-NP-exposed trophoblasts. Phenyl-N-t-butylnitrone (PBN), a free radical scavenger, inhibited 1-NP-induced genotoxic stress and trophoblast cycle arrest. Animal experiment showed that N-acetylcysteine (NAC), an antioxidant, rescued 1-NP-induced placental proliferation inhibition and fetal IUGR in mice. These results provide evidence that reactive oxygen species (ROS)-mediated cellular genotoxic stress partially contributes to 1-NP-induced placental proliferation inhibition and fetal IUGR.


Assuntos
Retardo do Crescimento Fetal , Mutagênicos/toxicidade , Pirenos/toxicidade , Trofoblastos , Animais , Ciclo Celular/efeitos dos fármacos , Dano ao DNA , Feminino , Humanos , Camundongos , Placenta , Gravidez , Pirenos/química , Espécies Reativas de Oxigênio/metabolismo
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