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1.
J Pregnancy ; 2022: 3922368, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35494491

RESUMO

Preeclampsia is a serious pregnancy disorder which in extreme cases may lead to maternal and fetal injury or death. Preexisting conditions which increase oxidative stress, e.g., hypertension and diabetes, increase the mother's risk to develop preeclampsia. Previously, we established that when the extracellular matrix is exposed to oxidative stress, trophoblast function is impaired, and this may lead to improper placentation. We investigated how the oxidative ECM present in preeclampsia alters the behavior of first trimester extravillous trophoblasts. We demonstrate elevated levels of advanced glycation end products (AGE) and lipid oxidation end product 4-hydroxynonenal in preeclamptic ECM (28%, and 32% increase vs control, respectively) accompanied with 35% and 82% more 3-chlorotyrosine and 3-nitrotyrosine vs control, respectively. Furthermore, we hypothesized that 670 nm phototherapy, which has antioxidant properties, reverses the observed trophoblast dysfunction as depicted in the improved migration and reduction in apoptosis. Since NO is critical for placentation, we examined eNOS activity in preeclamptic placentas compared to healthy ones and found no differences; however, 670 nm light treatment triggered enhanced NO availability presumably by using alternative NO sources. Light exposure decreased apoptosis and restored trophoblast migration to levels in trophoblasts cultured on preeclamptic ECM. Moreover, 670 nm irradiation restored expression of Transforming Growth Factor (TGFß) and Placental Growth Factor (PLGF) to levels observed in trophoblasts cultured on healthy placental ECM. We conclude the application of 670 nm light can successfully mitigate the damaged placental microenvironment of late onset preeclampsia as depicted by the restored trophoblast behavior.


Assuntos
Pré-Eclâmpsia , Trofoblastos , Matriz Extracelular/metabolismo , Feminino , Humanos , Placenta/metabolismo , Fator de Crescimento Placentário , Placentação , Pré-Eclâmpsia/metabolismo , Gravidez , Trofoblastos/metabolismo
2.
Sci Rep ; 12(1): 7348, 2022 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-35513694

RESUMO

Zika virus (ZIKV) infection at the maternal-placental interface is associated with adverse pregnancy outcomes including fetal demise and pregnancy loss. To determine how infection impacts placental trophoblasts, we utilized rhesus macaque trophoblast stem cells (TSC) that can be differentiated into early gestation syncytiotrophoblasts (ST) and extravillous trophoblasts (EVT). TSCs and STs, but not EVTs, were highly permissive to productive infection with ZIKV strain DAK AR 41524. The impact of ZIKV on the cellular transcriptome showed that infection of TSCs and STs increased expression of immune related genes, including those involved in type I and type III interferon responses. ZIKV exposure altered extracellular vesicle (EV) mRNA, miRNA and protein cargo, including ZIKV proteins, regardless of productive infection. These findings suggest that early gestation macaque TSCs and STs are permissive to ZIKV infection, and that EV analysis may provide a foundation for identifying non-invasive biomarkers of placental infection in a highly translational model.


Assuntos
Vesículas Extracelulares , Complicações Infecciosas na Gravidez , Infecção por Zika virus , Zika virus , Animais , Vesículas Extracelulares/genética , Feminino , Expressão Gênica , Humanos , Macaca mulatta , Placenta/metabolismo , Gravidez , Trofoblastos/metabolismo , Zika virus/fisiologia
3.
Vet Res ; 53(1): 33, 2022 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-35505413

RESUMO

Autophagy has been demonstrated to play important roles in the infection and pathogenesis of many viruses. We previously found that porcine parvovirus (PPV) infection can induce autophagy in porcine placental trophoblast cells (PTCs), but its underlying mechanism has not yet been fully revealed. In this study, we showed that PPV infection inhibited the activation of mTORC1 and promoted the expression of Beclin 1 and LC3II in PTCs. Treatment with a mTOR activator inhibited the expression of Beclin 1 and LC3II, as well as autophagy formation, and reduced viral replication in PPV-infected PTCs. Furthermore, we found that inhibition of AMPK expression, but not the inhibition of PI3K/Akt, p53, or MAPK/ERK1/2 pathway activation, can significantly increase mTOR phosphorylation in PPV-infected PTCs. Then, we found that the regulation of mTOR phosphorylation by AMPK was mediated by Raptor. AMPK expression knockout inhibited the activation of Raptor, decreased the expression of Beclin 1 and LC3II, suppressed the formation of autophagosomes, and reduced viral replication during PPV infection. Together, our results showed that PPV infection induces autophagy to promote viral replication by inhibiting the activation of mTORC1 through activation of the AMPK/Raptor pathway. These findings provide information to understand the molecular mechanisms of PPV-induced autophagy.


Assuntos
Infecções por Parvoviridae , Parvovirus Suíno , Aves Predatórias , Doenças dos Suínos , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Autofagia , Proteína Beclina-1 , Feminino , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Infecções por Parvoviridae/veterinária , Fosfatidilinositol 3-Quinases/metabolismo , Placenta , Gravidez , Aves Predatórias/metabolismo , Transdução de Sinais , Suínos , Serina-Treonina Quinases TOR/metabolismo , Trofoblastos/metabolismo , Replicação Viral
4.
Nan Fang Yi Ke Da Xue Xue Bao ; 42(4): 610-617, 2022 Apr 20.
Artigo em Chinês | MEDLINE | ID: mdl-35527499

RESUMO

OBJECTIVE: To investigate the expression of Talin1 in the fallopian tube and chorionic villi in patients with tubal pregnancy and its role in regulating invasion and migration of trophoblasts. METHODS: Immunohistochemistry and Western blotting were used to detect the localization and expression level of Talin1 in the fallopian tube and chorionic villi in patients with tubal pregnancy and in women with normal pregnancy. In the cell experiment, HTR-8/SVneo cells was transfected with Talin1 siRNA and the changes in cell invasion and migration were assessed using scratch assay and Transwell assay. The expressions of MMP-2, MMP-9, N-cadherin and Snail in the transfected cells were detected by qRT-PCR and Western blotting. RESULTS: Positive expression of Talin1 was detected in both normal fallopian tube tissues and tissues from women tubal pregnancy, and its expression was localized mainly in the cytoplasm of cilia cells. The expression level of Talin1 was significantly higher in both the fallopian tube and chorionic villi in women with tubal pregnancy than in normal fallopian tube and chorionic villi samples (P < 0.01). In HTR-8/SVneo cells, transfection with Talin1 siRNA significantly inhibited cell invasion (P < 0.01) and migration (P < 0.05), down-regulated the expression of N-cadherin, MMP-2 and Snail (P < 0.05), and up-regulated the expression of MMP-9 in the cells (P < 0.05). CONCLUSION: The expression of Talin1 in the fallopian tube and chorionic villi is significantly increased in women with tubal pregnancy, suggesting the association of Talin1-regulated trophoblast cell invasion with the occurrence of tubal pregnancy.


Assuntos
Gravidez Tubária , Trofoblastos , Caderinas/metabolismo , Movimento Celular , Vilosidades Coriônicas/metabolismo , Tubas Uterinas/metabolismo , Feminino , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Gravidez , Gravidez Tubária/metabolismo , RNA Interferente Pequeno/metabolismo , Trofoblastos/metabolismo
5.
Cell Stem Cell ; 29(5): 810-825.e8, 2022 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-35523141

RESUMO

Trophoblast organoids derived from placental villi provide a 3D model system of human placental development, but access to first-trimester tissues is limited. Here, we report that trophoblast stem cells isolated from naive human pluripotent stem cells (hPSCs) can efficiently self-organize into 3D stem-cell-derived trophoblast organoids (SC-TOs) with a villous architecture similar to primary trophoblast organoids. Single-cell transcriptome analysis reveals the presence of distinct cytotrophoblast and syncytiotrophoblast clusters and a small cluster of extravillous trophoblasts, which closely correspond to trophoblast identities in the post-implantation embryo. These organoid cultures display clonal X chromosome inactivation patterns previously described in the human placenta. We further demonstrate that SC-TOs exhibit selective vulnerability to emerging pathogens (SARS-CoV-2 and Zika virus), which correlates with expression levels of their respective entry factors. The generation of trophoblast organoids from naive hPSCs provides an accessible 3D model system of the developing placenta and its susceptibility to emerging pathogens.


Assuntos
COVID-19 , Células-Tronco Pluripotentes , Infecção por Zika virus , Zika virus , Diferenciação Celular , Feminino , Humanos , Organoides , Placenta/metabolismo , Placentação , Células-Tronco Pluripotentes/metabolismo , Gravidez , SARS-CoV-2 , Trofoblastos/metabolismo , Infecção por Zika virus/metabolismo
6.
Medicine (Baltimore) ; 101(13): e29130, 2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-35421066

RESUMO

ABSTRACT: To study the relationship between miR-148a and preeclampsia (PE), and clarify that miR-148a can regulate the endoplasmic reticulum stress (ERS) of placental trophoblasts by targeting the ERS protein X box binding protein 1 (XBP1).Fifty patients with hypertension during pregnancy, patients with mild PE, patients with severe PE, and normal pregnant women were selected, and their placental tissues were collected. RT-polymerase chain reaction was used to detect the expression of miR-148a in placental tissues, and Western blot was used to detect XBP1 in placental tissues. Compare the expression differences of miR-148a and XBP1 in each group, and analyze the correlation between the expressions of the two.Compared with the Neg-miR group, MTT experiment result in pre-miR-148a group was decreased. MTT experiment result in anti-miR-148a group was increased. Cell cycle test result in pre-miR-148a group [G1 (%)] was increased. Cell cycle test result in anti-miR-148a group [S (%)] was increased. Apoptosis test result in pre-miR-148a group [early apoptotic cells (%), late apoptotic cells (%)] was increased. Apoptosis test result in anti-miR-148a group [early apoptotic cells (%), late apoptotic cells (%)] was decreased. XBP1 expression result in pre-miR-148a group was increased. XBP1 expression result in anti-miR-148a group was decreased. Compared with the normal population, XBP1 is expressed in hypertension, mild eclampsia, severe eclampsia increased. GRP78, CHOP, and caspase-12 expression result in pre-miR-148a group was increased. GRP78, CHOP, and caspase-12 expression result in anti-miR-148a group was decreased.miR-148a can regulate the ERS-mediated apoptosis by targeting XBP1, thereby intervening in the occurrence and development of PE.


Assuntos
Eclampsia , Hipertensão , MicroRNAs , Pré-Eclâmpsia , Antagomirs/metabolismo , Apoptose/genética , Caspase 12/metabolismo , Movimento Celular , Estresse do Retículo Endoplasmático/genética , Feminino , Humanos , Hipertensão/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Placenta/metabolismo , Pré-Eclâmpsia/genética , Pré-Eclâmpsia/metabolismo , Gravidez , Trofoblastos/metabolismo , Proteína 1 de Ligação a X-Box/genética , Proteína 1 de Ligação a X-Box/metabolismo
7.
J Transl Med ; 20(1): 180, 2022 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-35449053

RESUMO

BACKGROUND: Insulin resistance (IR) during gestational diabetes mellitus (GDM) has been linked to dysregulated insulin-PI3K/Akt pathway. A defective insulin-PI3K/Akt pathway and dysregulated circular RNA (circRNA) levels have been observed in the placentas of patients with GDM; however, the mechanisms underlying this association remain unclear. METHODS: circRNAs potentially associated with GDM were selected through bioinformatics analysis and initially identified by quantitative real-time PCR (qPCR) in 9 GDM patients and 9 healthy controls, of which circMAP3K4 was further validated in additional 84 samples by qPCR. circMAP3K4 identity and localization were verified. Pearson correlation analysis was applied to evaluate the correlation between circMAP3K4 expression in the placental tissues of GDM patients and IR-related indicators. An IR model of trophoblasts was constructed using glucosamine. Interactions between miR-6795-5p and circMAP3K4 or PTPN1 were confirmed using a dual-luciferase reporter assay. The circMAP3K4/miR-6795-5p/PTPN1 axis and key markers in the insulin-PI3K/Akt pathway in placentas and trophoblasts were evaluated through qRT-PCR, immunofluorescence, and western blotting. The role of circMAP3K4 in glucose metabolism and cell growth in trophoblasts was determined using the glucose uptake and CCK8 assay, respectively. RESULTS: circMAP3K4 was highly expressed in the placentas of patients with GDM and the IR trophoblast model; this was associated with a dysregulated insulin-PI3K/Akt pathway. circMAP3K4 in the placentas of GDM patients was positively correlated with weight gain during pregnancy and time-glucose area under the curve of OGTT. circMAP3K4 and PTPN1 could both bind to miR-6795-5p. miR-6795-5p and PTPN1 were downregulated and upregulated, respectively, in the placentas of GDM patients and the IR trophoblast model. circMAP3K4 silencing or miR-6795-5p overexpression partially reversed the decrease in glucose uptake, inhibition in cell growth, and downregulated IRS1 and Akt phosphorylation in IR-trophoblasts; this restoration was reversed upon co-transfection with an miR-6795-5p inhibitor or PTPN1. CONCLUSION: circMAP3K4 could suppress the insulin-PI3K/Akt signaling pathway via miR-6795-5p/PTPN1 axis, probably contributing to GDM-related IR.


Assuntos
Diabetes Gestacional , Resistência à Insulina , MicroRNAs , Diabetes Gestacional/genética , Feminino , Glucose/metabolismo , Humanos , Insulina/metabolismo , Resistência à Insulina/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Placenta/metabolismo , Gravidez , Proteína Tirosina Fosfatase não Receptora Tipo 1/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Circular/genética , Trofoblastos/metabolismo
8.
Ecotoxicol Environ Saf ; 236: 113508, 2022 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-35427876

RESUMO

Perfluorooctanoate acid (PFOA) is a highly persistent and widespread chemical in the environment. PFOA serum levels in pregnant women are positively associated with an increased risk of placenta-related disorders. However, the mechanism of PFOA cytotoxicity involved in placental cells and cellular responses such as ER stress remains poorly understood. In this study, we studied the cellular toxicity of PFOA with a focus on proliferation and apoptosis in a human placental trophoblast cell line. Cell viability, number, apoptosis, stress response, activation of the involved signaling pathways were assessed. Our results showed PFOA affected cell viability, proliferation and also resulted in apoptosis. Besides, both pro-proliferation and pro-apoptosis effects were attenuated by endoplasmic reticulum (ER) stress inhibitors. Further experiments demonstrated that two different signaling pathways were activated by PFOA-induced ER stress and involved in PFOA toxicity: the reactive oxygen species (ROS)-dependent ERK signaling triggered trophoblast proliferation, while the ATF4-dependent C/EBP homologous protein (CHOP) signaling was the trigger of apoptosis. We conclude that PFOA-induced ER stress is the trigger of proliferation and apoptosis of trophoblast via ROS or UPR signaling pathway, which leads to the altered balance critical to the normal development and function of the placenta.


Assuntos
Placenta , Trofoblastos , Apoptose , Caprilatos , Proliferação de Células , Estresse do Retículo Endoplasmático , Feminino , Fluorcarbonetos , Humanos , Placenta/metabolismo , Gravidez , Espécies Reativas de Oxigênio/metabolismo , Trofoblastos/metabolismo
9.
Biomed Res Int ; 2022: 4769790, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35434129

RESUMO

Introduction: Controlling the invasive activity of trophoblastic tissue has not been elucidated. In the accreta placenta, the invasion of placental tissue is directly related to the expression of CRIPTO-1 at the maternal-fetal interface. The aim of this study is to evaluate if the expression of the CRIPTO-1 is related to different degrees of trophoblast invasion into the tube wall in ampullary pregnancy. Methods: Prospective study with 21 patients with ampullary tubal pregnancy undergoing salpingectomy. Anatomopathological evaluation determined the degree of invasion of trophoblast tissues into the tubal wall and grouped the samples into invasive degrees I, II, or III. The groups were compared for tissue expression of CRIPTO-1 using the Kruskal-Wallis nonparametric test. p values lower than 0.05 were considered significant. Results: Quantitative expression of CRIPTO-1 differed in each of the three groups of trophoblast invasion in the tubal wall in ampullary pregnancies (p < 0.001). There is a difference between groups when grade I + grade II versus grade III (p < 0.001) and grade I versus grade II + grade III (p < 0.001). The tissue expression of CRIPTO-1 in ectopic trophoblasts showed that deeper invasion of the tubal wall was associated with stronger expression than in shallow invasion (p < 0.001). Discussion. In ampullary pregnancies, the depth of penetration of trophoblast tissue in the tubal wall is related to CRIPTO-1 tissue expression.


Assuntos
Gravidez Tubária , Trofoblastos , Tubas Uterinas/metabolismo , Feminino , Proteínas Ligadas por GPI , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Proteínas de Neoplasias , Placenta/metabolismo , Gravidez , Gravidez Tubária/metabolismo , Gravidez Tubária/patologia , Estudos Prospectivos , Trofoblastos/metabolismo
10.
Sci Rep ; 12(1): 6374, 2022 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-35430618

RESUMO

Fetal growth restriction (FGR) is a common obstetric disease, which is harmful to the pregnant women and fetuses. It has many influencing factors, but the specific etiology is not clear. MiRNA plays an important role in the fetal growth and development. In this article, we use TaqMan Low-Density Array to screen and analyze the differently expressed miRNAs in FGR-affected placenta (n = 40) and the normal placenta (n = 40). A total of 139 abnormally expressed miRNAs in the FGR-affected placenta were identified, and miR-1227-3p was the most highly downregulated miRNA. Importantly, miR-1227-3p may promote the proliferation in HTR-8/SVneo cells, while inhibited the apoptosis of HTR-8/SVneo cells. DAVID was used to analyze the pathway enrichment of target genes of miR-1227-3p to predict its mechanism of action. Furthermore, the putative targets of miR-1227-3p were predicted using the TargetScan, PicTar, DIANA LAB, and miRWalk database. The potential expression of target genes of miR-1227-3p, including PRKAB2, AKT1, PIK3R3, and MKNK1 were significantly increased in FGR-affected placenta. Taken together, miR-1227-3p may participate in the development of FGR via regulating trophoblast cell proliferation and apoptosis by targeting genes involved in the insulin pathway. MiR-1227-3p may have a potential clinical value in the prevention and treatment of FGR, we need to study further to prove its value in the future.


Assuntos
Retardo do Crescimento Fetal , MicroRNAs , Trofoblastos , Apoptose/genética , Proliferação de Células/genética , Feminino , Retardo do Crescimento Fetal/genética , Retardo do Crescimento Fetal/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Placenta/metabolismo , Gravidez , Trofoblastos/metabolismo
11.
Proc Natl Acad Sci U S A ; 119(18): e2200582119, 2022 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-35476530

RESUMO

SignificancePlasma membranes are composed of a lipid bilayer in which phosphatidylserine (PtdSer) is confined to the inner leaflet by the action of flippase that translocates PtdSer from the outer to inner leaflets. Two P4-ATPases (ATP11A and ATP11C) work as flippase at plasma membranes. Here, we report that the mouse placenta expresses only ATP11A, and Atp11a-deficient mouse embryos die during embryogenesis due to inefficient formation of syncytiotrophoblasts in the placental labyrinth. The flippase-null mutation inactivates human choriocarcinoma BeWo cells to translocate PtdSer into the inner leaflet and undergo cell fusion. These findings highlight the importance of flippase to regulate the distribution of phospholipids for cell fusion, at least in trophoblast fusion.


Assuntos
Placenta , Trofoblastos , Transportador 1 de Cassete de Ligação de ATP , Adenosina Trifosfatases/metabolismo , Animais , Membrana Celular/metabolismo , Feminino , Camundongos , Fosfatidilserinas/metabolismo , Placenta/metabolismo , Gravidez , Trofoblastos/metabolismo
12.
Taiwan J Obstet Gynecol ; 61(2): 255-264, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35361385

RESUMO

OBJECTIVE: Abnormal expression of long non-coding RNAs (lncRNAs) is critical in preeclampsia (PE) pathogenesis. The current study explored the function of non-coding RNA activated by DNA damage (NORAD) in the progression of PE. MATERIALS AND METHODS: Quantitative real-time PCR was utilized to determine the expression of NORAD, microRNA (miR)-202-5p, and fragile X-related gene 1 (FXR1) in PE and normal placenta tissues. Cell viability was determined using 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-h-tetrazolium bromide assay, and cell migration and invasion were assessed using the Transwell assay. Target relationships were confirmed with the dual-luciferase reporter assay. Western blotting was performed to determine the protein level of FXR1. RESULTS: NORAD expression was markedly reduced in PE placenta. NORAD over-expression enhanced the viability, migration, and invasion of HTR-8/SVneo cells. miR-202-5p was a target and was negatively regulated by NORAD. Down-regulation of miR-202-5p promoted the viability, migration, and invasion of HTR-8/SVneo cells. miR-202-5p inversely regulated FXR1 expression by targeting the 3'UTR of FXR1. Both miR-202-5p up-regulation and FXR1 knockdown reversed the NORAD over-expression-induced enhancement in the viability, migration, and invasion of HTR-8/SVneo cells. CONCLUSION: Collectively, the results revealed that NORAD over-expression promoted trophoblast viability, invasion, and migration by regulating the miR-202-5p/FXR1 axis. These findings clarify PE pathogenesis and will inform the discovery of new targets for PE treatment.


Assuntos
MicroRNAs , Pré-Eclâmpsia , RNA Longo não Codificante , Proteínas de Ligação a RNA , Sobrevivência Celular/genética , Feminino , Humanos , MicroRNAs/genética , Pré-Eclâmpsia/genética , Pré-Eclâmpsia/metabolismo , Gravidez , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Trofoblastos/metabolismo
13.
Mol Hum Reprod ; 28(5)2022 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-35451485

RESUMO

Epigenetic changes alter the expression of genes at both pre- and post-transcriptional levels without changing their DNA sequence. Accumulating evidence suggests that such changes can modify cellular behavior and characteristics required during development and in response to various extracellular stimuli. Trophoblast cells develop from the outermost trophectoderm layer of the blastocyst and undergo many phenotypic changes as the placenta develops. One such phenotypic change is differentiation of the epithelial natured cytotrophoblasts into the mesenchymal natured extravillous trophoblasts. The extravillous trophoblasts are primarily responsible for invading into the maternal decidua and thus establishing connection with the maternal spiral arteries. Any dysregulation of this process can have adverse effects on the pregnancy outcome. Hence, tight regulation of this epithelial-mesenchymal transition (EMT) is critical for successful pregnancy. This review summarizes the recent research on the epigenetic regulation of the EMT occurring in the trophoblast cells during placental development. The functional significance of chemical modifications of DNA and histone, which regulate transcription, as well as non-coding RNAs, which control gene expression post-transcriptionally, is discussed in relation to trophoblast biology.


Assuntos
Transição Epitelial-Mesenquimal , Trofoblastos , Epigênese Genética , Transição Epitelial-Mesenquimal/genética , Feminino , Humanos , Placenta/metabolismo , Placentação/genética , Gravidez , Trofoblastos/metabolismo
14.
Placenta ; 121: 164-172, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35364512

RESUMO

INTRODUCTION: The etiology of approximately half of patients with recurrent spontaneous abortion (RSA) has yet to be established. Granulocyte-colony stimulating factor (G-CSF) exerts a protective effect on pregnancy and its absence may lead to pregnancy failure. However, the effects and mechanisms of G-CSF activities have not been fully explored. Therefore, we aimed at evaluating whether a loss of G-CSF induces RSA by affecting cell communication at the maternal-foetal interface. METHODS: Villous and decidual tissues were obtained from participants and expression levels of G-CSF determined by qRT-PCR, Western blot and immunohistochemistry. G-CSF levels in trophoblasts were downregulated by siRNA. Exosomes were extracted from trophoblasts and co-cultured with macrophages. Molecular expression levels of key genes were determined by qRT-PCR and Western blot. Migration and proliferation of cells were evaluated by transwell and CCK8 assays. The RSA mice models were intraperitoneally administered with G-CSF to assess pregnancy outcomes and expression profiles of G-CSF as well as its receptor at the mother-foetal interface. RESULTS: Relative to the decidua, G-CSF was highly expressed in the villus, and expression levels were low in RSA tissues compared to normal tissues. Down-regulation of G-CSF in the trophoblast cell line (HTR-8/SVneo) by siRNA was associated with a decrease in cell activities. Trophoblast-derived exosomes inhibited the activation of the macrophage cell line (RAW264.7), whereas G-CSF free exosomes had no effects on macrophage activation. Intraperitoneal administration of G-CSF improved pregnancy outcomes in RSA mice and increased the amounts of G-CSF at the maternal-foetal interface. DISCUSSION: G-CSF levels were downregulated in villi of RSA patients. The absence of G-CSF impaired the proliferation as well as migration capacities of trophoblasts, and weakened the suppression of trophoblasts against macrophages. This implies that suppressed G-CSF levels may be a key factor in RSA occurrence. G-CSF decreased the rate of abortion in RSA mice, thus, it could be a treatment option for RSA patients.


Assuntos
Aborto Habitual , Aborto Espontâneo , Aborto Habitual/metabolismo , Aborto Espontâneo/metabolismo , Animais , Movimento Celular , Feminino , Fator Estimulador de Colônias de Granulócitos/metabolismo , Humanos , Macrófagos/metabolismo , Camundongos , Gravidez , RNA Interferente Pequeno , Trofoblastos/metabolismo
15.
Mol Cell Endocrinol ; 549: 111642, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35395343

RESUMO

BACKGROUND: A supply of maternal thyroid hormone (thyroxine, T4) is essential for normal human fetal development. Human placental trophoblasts synthesize, secrete and take up the T4 binding protein transthyretin, providing a route for maternal T4 to enter the placenta. Transthyretin is also involved in T4 transport in other tissues such as the brain choroid plexus. Nicotine alters transthyretin synthesis and function in rat choroid plexus. If nicotine influences trophoblast turnover of transthyretin, then it may directly affect placental transfer of T4 to the developing fetus and contribute to the negative impacts of smoking on fetal growth, development and placental function. METHODS: The effect of nicotine on trophoblast uptake of Alexa-labelled transthyretin was measured using live cell imaging. The effect of nicotine on protein expression was measured by western blotting. Interactions between transthyretin, T4 and nicotine were investigated using chemical cross-linking techniques and molecular dynamic simulations. RESULTS: Nicotine blocks uptake of transthyretin-T4 by human placental trophoblast cells. Nicotine reduces the expression of the trophoblast scavenger receptor class B type 1 (SR-B1) that plays a role in transthyretin-T4 uptake. Molecular dynamic modelling suggests that when T4 is bound to transthyretin, nicotine binding increases tetramer stability, reducing the ability of the transthyretin-T4 complex to enter trophoblast cells. CONCLUSION: Our data suggest that nicotine exposure during pregnancy reduces transplacental transport of transthyretin and T4 to the placenta and developing fetus. This may contribute to the negative effects of smoking on fetal growth, development and pregnancy viability.


Assuntos
Tiroxina , Trofoblastos , Animais , Feminino , Nicotina/farmacologia , Placenta/metabolismo , Pré-Albumina/metabolismo , Gravidez , Ratos , Fumar , Tiroxina/metabolismo , Tiroxina/farmacologia , Trofoblastos/metabolismo
16.
Ecotoxicol Environ Saf ; 237: 113564, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35483139

RESUMO

Human trophoblast cell apoptosis may induce miscarriage. Trophoblast cells are sensitive to environmental BaP-7,8-dihydrodiol-9,10-epoxide (BPDE). However, how BPDE induces human trophoblast cell apoptosis is still largely elusive. In this work, we used BPDE-treated human trophoblast cells and villous tissues collected from recurrent miscarriage and health control groups to explore the underlying mechanism of BPDE-induced human trophoblast cell apoptosis. Continued with our recent work, we found that lncRNA HZ01 (lnc-HZ01) could induce human trophoblast cell apoptosis. In mechanism, lnc-HZ01 up-regulated p53 expression level by suppressing its MDM2-mediated proteasomal degradation. Meanwhile, we found that p53 acted as lnc-HZ01 transcription factor and promoted lnc-HZ01 transcription. Thus, lnc-HZ01 and p53 composed a positive feedback loop in human trophoblast cells. In normal trophoblast cells, relatively low levels of lnc-HZ01 and p53 suppressed p53/caspase-3 apoptosis pathway, giving normal pregnancy. Upon BPDE exposure, BPDE up-regulated the expression levels of lnc-HZ01 and p53, triggered this positive feedback loop, activated the p53/caspase-3 apoptosis pathway, and then induced miscarriage. Collectively, we discovered new mechanism by which lnc-HZ01 regulated BPDE-induced human trophoblast cell apoptosis, providing scientific basis for the diagnosis and treatment of unexplained recurrent miscarriage.


Assuntos
Aborto Habitual , RNA Longo não Codificante , 7,8-Di-Hidro-7,8-Di-Hidroxibenzo(a)pireno 9,10-óxido/metabolismo , 7,8-Di-Hidro-7,8-Di-Hidroxibenzo(a)pireno 9,10-óxido/toxicidade , Aborto Habitual/induzido quimicamente , Aborto Habitual/metabolismo , Apoptose , Caspase 3/metabolismo , Retroalimentação , Feminino , Humanos , Gravidez , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Trofoblastos/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo
17.
Cells ; 11(7)2022 03 29.
Artigo em Inglês | MEDLINE | ID: mdl-35406722

RESUMO

Spiral-artery (SA) remodeling is a fundamental process during pregnancy that involves the action of cells of the initial vessel, such as vascular smooth-muscle cells (VSMCs) and endothelial cells, but also maternal immune cells and fetal extravillous trophoblast cells (EVTs). Mast cells (MCs), and specifically chymase-expressing cells, have been identified as key to a sufficient SA-remodeling process in vivo. However, the mechanisms are still unclear. The purpose of this study is to evaluate the effects of the MC line HMC-1 and recombinant human chymase (rhuCMA1) on human primary uterine vascular smooth-muscle cells (HUtSMCs), a human trophoblast cell line (HTR8/SV-neo), and human umbilical-vein endothelial cells (HUVEC) in vitro. Both HMC-1 and rhuCMA1 stimulated migration, proliferation, and changed protein expression in HUtSMCs. HMC-1 increased proliferation, migration, and changed gene expression of HTR8/SVneo cells, while rhuCMA treatment led to increased migration and decreased expression of tissue inhibitors of matrix metalloproteinases. Additionally, rhuCMA1 enhanced endothelial-cell-tube formation. Collectively, we identified possible mechanisms by which MCs/rhuCMA1 promote SA remodeling. Our findings are relevant to the understanding of this crucial step in pregnancy and thus of the dysregulated pathways that can lead to pregnancy complications such as fetal growth restriction and preeclampsia.


Assuntos
Mastócitos , Trofoblastos , Quimases/metabolismo , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Miócitos de Músculo Liso/metabolismo , Fenótipo , Gravidez , Trofoblastos/metabolismo
18.
Oxid Med Cell Longev ; 2022: 7856290, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35464758

RESUMO

Objectives: This study is aimed at investigating the role of PIWIL1/piRNA in the development of preeclampsia. Methods: High-throughput sequencing was performed in 5 preeclampsia and 5 normal placentas to get a piRNA expression profile. WGCNA network was constructed to find hub piRNAs. Through target gene prediction and protein interaction network analysis, we found the potential relationship between the key genes and PIWIL1. Subsequently, we detected the expression of PIWIL1 in 35 preeclampsia and 29 normal placental tissues. Overexpression and inhibition of PIWIL1 in HTR-8/SVneo trophoblast cells were achieved by transfecting an overexpression vector and siRNAs, respectively. Cell proliferation, apoptosis, and invasion were assessed using CCK-8, flow cytometric, and transwell assays, respectively. Results: It was found that a total of three piRNAs were upregulated in preeclampsia (pir-hsa-1256314, uniq_271431, and uniq_277797). And two target genes with the highest connectivity (FXR1 and DDX6) both pointed to PIWIL1. PIWIL1 expression was significantly lower in preeclampsia. In vitro studies linked PIWIL1 expression to trophoblast overgrowth. Overexpression of PIWIL1 remarkably promoted cell proliferation and invasion and inhibited apoptosis of HTR-8/SVneo cells and vice versa. Conclusions: PIWIL1/piRNA may be involved in the pathogenesis of preeclampsia by inhibiting the proliferation and invasion and promoting the apoptosis of placental trophoblasts. This study was registered with the China Clinical Trials Registry (http://www.clinicaltrials.gov): registration number ChiCTR1900027479.


Assuntos
Pré-Eclâmpsia , Trofoblastos , Apoptose/genética , Proteínas Argonauta/genética , Proteínas Argonauta/metabolismo , Movimento Celular , Proliferação de Células/genética , Feminino , Humanos , Placenta/metabolismo , Pré-Eclâmpsia/metabolismo , Gravidez , RNA Interferente Pequeno/metabolismo , Proteínas de Ligação a RNA/metabolismo , Trofoblastos/metabolismo
19.
Front Endocrinol (Lausanne) ; 13: 842587, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35299960

RESUMO

The acquisition of an endovascular trophoblast (enEVT) phenotype is essential for normal placental development and healthy pregnancy. MicroRNAs (miRNAs) are small noncoding RNAs that play critical roles in regulating gene expression. We have recently reported that miR-218-5p promotes enEVT differentiation and spiral artery remodeling in part by targeting transforming growth factor ß2 (TGFß2). We also identified IL1B, which encodes interleukin 1ß (IL1ß), as one of the most highly upregulated genes by miR-218-5p. In this study, we investigated how miR-218-5p regulates IL1B expression and IL1ß secretion and the potential role of IL1ß in enEVT differentiation. Using two cell lines derived from extravillous trophoblasts (EVTs), HTR-8/SVneo and Swan 71, we found that stable overexpression of miR-218-5p precursor, mir-218-1, or transient transfection of miR-218-5p mimic, significantly increased IL1B mRNA and IL1ß protein levels in cells and conditioned media. We also showed that miR-218-5p directly interacted with SMAD2 3'UTR and reduced SMAD2 at mRNA and protein levels. Knockdown of SMAD2 induced IL1B expression and attenuated the inhibitory effect of TGFß2 on IL1B expression. On the other hand, overexpression of SMAD2 reduced IL1ß levels and blocked the stimulatory effects of miR-218-5p on IL1B expression, trophoblast migration and endothelial-like network formation. In addition, treatment of trophoblasts with IL1ß induced the formation of endothelial-like networks and the expression of enEVT markers in a dose-dependent manner. These results suggest that miR-218-5p inhibits the TGFß/SMAD2 pathway to induce IL1ß and enEVT differentiation. Finally, low doses of IL1ß also inhibited the expression of miR-218-5p, suggesting the existence of a negative feedback regulatory loop. Taken together, our findings suggest a novel interactive miR-218-5p/TGFß/SMAD2/IL1ß signaling nexus that regulates enEVT differentiation.


Assuntos
MicroRNAs , Trofoblastos , Regiões 3' não Traduzidas , Feminino , Humanos , Interleucina-1beta/farmacologia , MicroRNAs/genética , MicroRNAs/metabolismo , Placenta/metabolismo , Gravidez , Proteína Smad2/genética , Trofoblastos/metabolismo
20.
Nat Commun ; 13(1): 1626, 2022 03 25.
Artigo em Inglês | MEDLINE | ID: mdl-35338152

RESUMO

The combination of EGF, CHIR99021, A83-01, SB431542, VPA, and Y27632 (EGF/CASVY) facilitates the derivation of trophoblast stem (TS) cells from human blastocysts and first-trimester, but not term, cytotrophoblasts. The mechanism underlying this chemical induction of TS cells remains elusive. Here we demonstrate that the induction efficiency of cytotrophoblast is determined by functional antagonism of the placental transcription factor GCM1 and the stemness regulator ΔNp63α. ΔNp63α reduces GCM1 transcriptional activity, whereas GCM1 inhibits ΔNp63α oligomerization and autoregulation. EGF/CASVY cocktail activates ΔNp63α, thereby partially inhibiting GCM1 activity and reverting term cytotrophoblasts into stem cells. By applying hypoxia condition, we can further reduce GCM1 activity and successfully induce term cytotrophoblasts into TS cells. Consequently, we identify mitochondrial creatine kinase 1 (CKMT1) as a key GCM1 target crucial for syncytiotrophoblast differentiation and reveal decreased CKMT1 expression in preeclampsia. Our study delineates the molecular underpinnings of trophoblast stemness and differentiation and an efficient method to establish TS cells from term placentas.


Assuntos
Fator de Crescimento Epidérmico , Trofoblastos , Diferenciação Celular , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Fator de Crescimento Epidérmico/metabolismo , Feminino , Humanos , Proteínas Nucleares/metabolismo , Placenta/metabolismo , Gravidez , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Trofoblastos/metabolismo , Proteínas Supressoras de Tumor
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