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1.
Cell Prolif ; 53(2): e12745, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31889361

RESUMO

OBJECTIVES: The transformation of cytotrophoblasts into mesenchymal-like extravillous trophoblasts is necessary for successful embryo implantation, and the inadequate transformation may cause abortion. Epiregulin, which is a new growth factor, plays important roles in the reproductive processes. The glycosylation of many proteins in reproduction processes is critical. Protein O-fucosyltransferase 1 (poFUT1) is the key enzyme for the biosynthesis of O-fucosylation on the specific glycoproteins. Urokinase-type plasminogen activator (uPA) contains O-fucosylated domain on Thr18 . However, the functions of epiregulin and poFUT1 in the trophoblast epithelial-mesenchymal transition (EMT) process, the regulatory mechanism of epiregulin on poFUT1 and the resulting O-fucosylated uPA remain unclear. MATERIALS AND METHODS: We employed ELISA and Western blot to detect serum levels of epiregulin and poFUT1 from non-pregnancy women, pregnancy women and abortion patients. Using two trophoblast cell lines and a mouse pregnancy model, we investigated the underlying mechanisms of epiregulin and poFUT1 in trophoblast EMT process. RESULTS: Serum levels of epiregulin and poFUT1 were higher in pregnant women compared with non-pregnant women, and their levels were significantly decreased in abortion patients compared with pregnant women. The results showed that epiregulin upregulated poFUT1 expression and increased O-fucosylation on uPA, which further activated the PI3K/Akt signalling pathway, facilitating EMT behaviour of trophoblast cells and embryo implantation in the mouse pregnant model. CONCLUSIONS: Level of epiregulin and poFUT1 is lower in abortion patients than early pregnancy women. Epiregulin promotes trophoblast EMT through O-fucosylation on uPA catalysed by poFUT1. Epiregulin and poFUT1 may be suggested as the potential diagnostic biomarkers and useful treatment targets for abortion.


Assuntos
Epirregulina/metabolismo , Transição Epitelial-Mesenquimal/fisiologia , Fucosiltransferases/metabolismo , Trofoblastos/metabolismo , Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Adulto , Animais , Linhagem Celular , Feminino , Glicosilação , Humanos , Camundongos , Fosfatidilinositol 3-Quinases/metabolismo , Gravidez , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/fisiologia , Regulação para Cima/fisiologia
2.
Biochemistry (Mosc) ; 84(10): 1186-1196, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31694514

RESUMO

Studies of interactions between natural killer (NK) cells and trophoblasts and identification of conditions for the NK cells to perform their cytotoxic function are of fundamental and practical importance for understanding their role in the development of pathological processes and complications during pregnancy. In this study, we examined changes in the content of caspases and studied activation of these enzymes in Jeg-3 trophoblasts in various models of their coculturing with NK-92 cells and demonstrated the necessity of direct contact between these cell populations for the activation of caspase-8 and caspase-3 in the trophoblasts. Contact coculturing of the two cell lines resulted in the appearance of the cytotoxic protein granzyme B in Jeg-3 cells that was accompanied by a decrease in the content of this enzyme in NK-92 cells. Distant coculturing of NK-92 and Jeg-3 cells did not trigger initiator and effector caspases characteristic for the apoptosis development in Jeg-3 cells. The observed decrease in the content of procaspases in the trophoblasts may be associated with alternative non-apoptotic functions of these enzymes.


Assuntos
Caspases/metabolismo , Técnicas de Cocultura , Células Matadoras Naturais/metabolismo , Modelos Biológicos , Trofoblastos/metabolismo , Linhagem Celular Tumoral , Humanos
3.
Medicine (Baltimore) ; 98(44): e17676, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31689783

RESUMO

BACKGROUND: This study aimed to clarify the change of the expression of placenta-specific 1 (PLAC1) in the placenta of preeclamptic women and to explore the regulatory effects on thophoblast by PLAC1. METHODS: Nineteen women with preeclampsia and 19 with normal pregnancies were recruited, and then we determined the expression of PLAC1 by immunohistochemistry (IHC) and Western blotting. To observe the effect of hypoxia on the expression of PLAC1, trophoblasts were cultured at the normoxia or hypoxia condition. Small interference of ribonucleic acid (siRNA) was used to silence PLAC1. The proliferation, migration and invasion of trophoblasts were evaluated with cell counting kit-8 and transwell analysis, and the apoptosis of trophoblast was evaluated by flow cytometry with FITC and PI staining. RESULTS: Placental PLAC1 expression was significantly decreased in severe preeclampsia compared with control (P < .001). The expression of PLAC1 in trophoblasts was significantly decreased after treated with low oxygen concentration (P = .018). PLAC1 siRNA significantly inhibited the proliferation (P < .001), the migration (P < .001) and invasion (P < .001) of trophoblasts, but increased the apoptosis (P = .004 for Swan-71; P = .031 for Jar). CONCLUSIONS: The expression of PLAC1 was declined in preeclampsia and this inhibited the function of trophoblast, suggesting PLAC1 may play a role in the development of preeclampsia.


Assuntos
Hipóxia/fisiopatologia , Placenta/metabolismo , Pré-Eclâmpsia/metabolismo , Proteínas da Gravidez/biossíntese , Trofoblastos/metabolismo , Adulto , Apoptose , Movimento Celular , Proliferação de Células , Feminino , Humanos , Imuno-Histoquímica , Gravidez , RNA Interferente Pequeno , Índice de Gravidade de Doença
4.
Adv Exp Med Biol ; 1141: 505-548, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31571173

RESUMO

The placenta is the only organ linking two different individuals, mother and fetus, termed as blood-placental barrier. The functions of the blood-placental barrier are to regulate material transfer between the maternal and fetal circulation. The main functional units are the chorionic villi within which fetal blood is separated by only three or four cell layers (placental membrane) from maternal blood in the surrounding intervillous space. A series of drug transporters such as P-glycoprotein (P-GP), breast cancer resistance protein (BCRP), multidrug resistance-associated proteins (MRP1, MRP2, MRP3, MRP4, and MRP5), organic anion-transporting polypeptides (OATP4A1, OATP1A2, OATP1B3, and OATP3A1), organic anion transporter 4 (OAT4), organic cation transporter 3 (OCT3), organic cation/carnitine transporters (OCTN1 and OCTN2), multidrug and toxin extrusion 1 (MATE1), and equilibrative nucleoside transporters (ENT1 and ENT2) have been demonstrated on the apical membrane of syncytiotrophoblast, some of which also expressed on the basolateral membrane of syncytiotrophoblast or fetal capillary endothelium. These transporters are involved in transport of most drugs in the placenta, in turn, affecting drug distribution in fetus. Moreover, expressions of these transporters in the placenta often vary along with the gestational ages and are also affected by pathophysiological factor. This chapter will mainly illustrate function and expression of these transporters in placentas, their contribution to drug distribution in fetus, and their clinical significance.


Assuntos
Regulação da Expressão Gênica , Proteínas de Membrana Transportadoras , Placenta , Feminino , Humanos , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Placenta/metabolismo , Gravidez , Distribuição Tecidual , Trofoblastos/metabolismo
5.
Nat Commun ; 10(1): 4749, 2019 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-31628347

RESUMO

Trophectoderm (TE) lineage development is pivotal for proper implantation, placentation, and healthy pregnancy. However, only a few TE-specific transcription factors (TFs) have been systematically characterized, hindering our understanding of the process. To elucidate regulatory mechanisms underlying TE development, here we map super-enhancers (SEs) in trophoblast stem cells (TSCs) as a model. We find both prominent TE-specific master TFs (Cdx2, Gata3, and Tead4), and >150 TFs that had not been previously implicated in TE lineage, that are SE-associated. Mapping targets of 27 SE-predicted TFs reveals a highly intertwined transcriptional regulatory circuitry. Intriguingly, SE-predicted TFs show 4 distinct expression patterns with dynamic alterations of their targets during TSC differentiation. Furthermore, depletion of a subset of TFs results in dysregulation of the markers for specialized cell types in placenta, suggesting a role during TE differentiation. Collectively, we characterize an expanded TE-specific regulatory network, providing a framework for understanding TE lineage development and placentation.


Assuntos
Células-Tronco Embrionárias/metabolismo , Elementos Facilitadores Genéticos/genética , Regulação da Expressão Gênica no Desenvolvimento , Redes Reguladoras de Genes , Trofoblastos/metabolismo , Animais , Diferenciação Celular/genética , Linhagem da Célula/genética , Embrião de Mamíferos/citologia , Embrião de Mamíferos/embriologia , Embrião de Mamíferos/metabolismo , Feminino , Perfilação da Expressão Gênica/métodos , Camundongos , Placentação/genética , Gravidez , Fatores de Transcrição/genética , Trofoblastos/citologia
6.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 50(3): 328-333, 2019 May.
Artigo em Chinês | MEDLINE | ID: mdl-31631598

RESUMO

Objective: To investigate the expression of miRNA-148b-3p and its target gene in the placenta between normal pregnant women and pregnant women with intrahepatic cholestasis of pregnancy (ICP) and to explore the possible mechanism of glucose metabolism of offspring with maternal cholestasis. Methods: There were 30 cases of normal pregnant women and 30 cases of pregnant women with ICP recruited in the study, all of whom underwent cesarean delivery from Mar. 2017 to Jan. 2018. Placenta tissues, maternal blood and cord blood were collected in each case. Maternal blood and cord blood were sent for biochemical detection. miRNA of placenta tissues was extracted and qRT-PCR was used to measure the expression of miR-148b-3p in the placenta. Normal HTR-8 cells were transfected with miR-148b-3p inhibitor/mimics wrapped with lipofectaine3000. qRT-PCR was used to measure the expression of miR-148b-3p, and Western blot was used to measure the expression of glucose transporter 1 (GLUT1) after transfection. Results: Maternal fasting blood glucose (FPG) and its fetal cord blood insulin levels in the ICP group were significantly higher than those of control. The expression of miR-148b-3p in the placenta of ICP group was lower than that of control group ( P<0.05). Compared with inhibitor control group, the expression of miR-148b-3p was decreased in HTR-8 cells transfected with miR-148b-3p inhibitor ( P<0.05), while the expression of GLUT1 was increased ( P<0.05). Compared with mimics control group, the expression of miR-148b-3p was increased in HTR-8 cells transfected with miR-148b-3p mimics ( P<0.05), while the expression of GLUT1 was decreased ( P<0.05). Conclusion: miR-148b-3p might participate in glucose metabolism of offspring with maternal cholestasis through the negative regulation of GLUT1 expression in placental trophoblast cells.


Assuntos
Colestase Intra-Hepática/genética , Transportador de Glucose Tipo 1/genética , Glucose/metabolismo , MicroRNAs/genética , Placenta/citologia , Complicações na Gravidez/genética , Trofoblastos/metabolismo , Estudos de Casos e Controles , Feminino , Humanos , Gravidez
7.
Life Sci ; 236: 116899, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31614145

RESUMO

AIMS: The aim of our study is to illustrate the role of amphiregulin in trophoblast invasiveness and underlying signal cascades. MAIN METHODS: An immortalized human early extravillous cell line, HTR-8/SVneo, was used for this investigation. Matrigel-transwell invasion assay was used for testing the effects of amphiregulin on cell invasiveness. MMP9 and MMP2 mRNA expression level and activity were measured using Rt-qPCR and zymographic analysis. Cell signals involved in the invasion process were verified using western blot and specific inhibitors. KEY FINDINGS: Our results showed that amphiregulin could promote HTR-8/SVneo cell invasiveness without interfering cell proliferation, and significantly upregulate the expression of MMP9 and TIMP-1 mRNAs as well as the ratio of MMP9/TIMP-1. Using specific inhibitors for MEK and PI3K signaling further indicated that, both ERK1/2 and Akt signal pathways were required for amphiregulin-induced cell invasiveness. The co-ordination between ERK1/2 and Akt signaling pathway was needed for the upregulation of MMP9 mRNA, while ERK1/2 was more essential for the upregulation of TIMP-1 mRNA. Meanwhile, we first put forward that the deficiency of amphiregulin expression in trophoblast might be compensated by the upregulation of epidermal growth factor receptor (EGFR) and heparin-binding EGF (HB-EGF) mRNA. SIGNIFICANCE: ERK1/2 and Akt signaling pathways mediate amphiregulin-induced upregulation of MMP9 mRNA and the MMP9/TIMP-1 ratio, which subsequently contribute to amphiregulin-promotion of HTR-8/SVneo cell invasion.


Assuntos
Anfirregulina/farmacologia , Metaloproteinase 9 da Matriz/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Trofoblastos/metabolismo , Trofoblastos/patologia , Movimento Celular , Proliferação de Células , Células Cultivadas , Feminino , Regulação da Expressão Gênica , Humanos , Metaloproteinase 9 da Matriz/genética , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/genética , Gravidez , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais , Inibidor Tecidual de Metaloproteinase-1/genética
8.
PLoS Biol ; 17(10): e3000187, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31596842

RESUMO

Multipotent trophoblasts undergo dynamic morphological movement and cellular differentiation after conceptus implantation to generate placenta. However, the mechanism controlling trophoblast development and differentiation during peri-implantation development in human remains elusive. In this study, we modeled human conceptus peri-implantation development from blastocyst to early postimplantation stages by using an in vitro coculture system and profiled the transcriptome of 476 individual trophoblast cells from these conceptuses. We revealed the genetic networks regulating peri-implantation trophoblast development. While determining when trophoblast differentiation happens, our bioinformatic analysis identified T-box transcription factor 3 (TBX3) as a key regulator for the differentiation of cytotrophoblast (CT) into syncytiotrophoblast (ST). The function of TBX3 in trophoblast differentiation is then validated by a loss-of-function experiment. In conclusion, our results provided a valuable resource to study the regulation of trophoblasts development and differentiation during human peri-implantation development.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Redes Reguladoras de Genes , Modelos Biológicos , Proteínas com Domínio T/genética , Transcriptoma , Trofoblastos/metabolismo , Técnicas de Cultura de Células , Diferenciação Celular , Biologia Computacional/métodos , Implantação do Embrião/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Análise de Célula Única , Proteínas com Domínio T/metabolismo , Trofoblastos/citologia , Zigoto
9.
Nat Commun ; 10(1): 4601, 2019 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-31601798

RESUMO

During pregnancy, trophoblast cells sustain the maternal-fetal tolerance via expressing and secreting various chemokines and cytokines. Our previous study revealed the expression of interleukin-35 (IL-35) in human first-trimester trophoblasts. Here we show that IL-35 is expressed in both human first-trimester primary trophoblast cells and a trophoblast cell line. Trophoblast cells inhibit the proliferation of human naive conventional T cells (Tconv cells) and convert suppressed Tconv cells into iTR35 in an IL-35-dependent manner. Mechanistically, trophoblast cell derived IL-35 mediates its function through phosphorylation of STAT1 and STAT3. In vivo studies confirm that mice with immunologically spontaneous abortion have lower levels of IL-35 and iTR35 cells at the maternal-fetal interface, and neutralizing anti-IL-35 mAb enhances abortion rates. Meanwhile, exogenous IL-35 induces iTR35 and prevents immunological abortion. Our findings thus suggest that trophoblast cells have a critical function in preserving maternal-fetal tolerance via secreting IL-35 during pregnancy.


Assuntos
Interleucinas/metabolismo , Troca Materno-Fetal/fisiologia , Linfócitos T Reguladores/imunologia , Trofoblastos/citologia , Animais , Proliferação de Células , Feminino , Humanos , Tolerância Imunológica , Interleucinas/sangue , Interleucinas/imunologia , Interleucinas/farmacologia , Masculino , Troca Materno-Fetal/imunologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos DBA , Placenta/citologia , Placenta/imunologia , Gravidez , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/farmacologia , Linfócitos T/citologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Linfócitos T Reguladores/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Trofoblastos/imunologia , Trofoblastos/metabolismo
10.
J Steroid Biochem Mol Biol ; 195: 105470, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31509772

RESUMO

Serotonin reuptake inhibitors (SRIs) are currently the main molecules prescribed to pregnant women that suffer from depression. Placental cells are exposed to SRIs via maternal blood, and we have previously shown that SRIs alter feto-placental steroidogenesis in an in vitro co-culture model. More specifically, serotonin (5-HT) regulates the estrogen biosynthetic enzyme aromatase (cytochrome P450 19; CYP19), which is disrupted by fluoxetine and its active metabolite norfluoxetine in BeWo choriocarcinoma cells. Based on molecular simulations, the present study illustrates that the SRIs fluoxetine, norfluoxetine, paroxetine, sertraline, citalopram and venlafaxine exhibit binding affinity for the active-site pocket of CYP19, suggesting potential competitive inhibition. Using BeWo cells and primary villous trophoblast cells isolated from normal term placentas, we compared the effects of the SRIs on CYP19 activity. We observed that paroxetine and sertraline induce aromatase activity in BeWo cells, while venlafaxine, fluoxetine, paroxetine and sertraline decrease aromatase activity in primary villous trophoblast. The effects of paroxetine and sertraline in primary villous trophoblasts were observed at the lower doses tested. We also showed that 5-HT and the 5-HT2A receptor agonist 2,5-dimethoxy-4-iodoamphetamine (DOI) induced CYP19 activity. An increase in phosphorylation of serine and tyrosine and a decrease in threonine phosphorylation of CYP19 was also associated with DOI treatment. Our results contribute to better understanding how 5-HT and SRIs interact with CYP19 and may affect estrogen production. Moreover, this study suggests that alteration of placental 5-HT levels due to depression and/or SRI treatment during pregnancy may be associated with disruption of placental estrogen production.


Assuntos
Aromatase/metabolismo , Placenta/efeitos dos fármacos , Inibidores de Captação de Serotonina/farmacologia , Serotonina/farmacologia , Células Cultivadas , Citalopram/farmacologia , Feminino , Fluoxetina/análogos & derivados , Fluoxetina/farmacologia , Humanos , Simulação de Acoplamento Molecular , Paroxetina/farmacologia , Placenta/metabolismo , Gravidez , Sertralina/farmacologia , Trofoblastos/efeitos dos fármacos , Trofoblastos/metabolismo , Cloridrato de Venlafaxina/farmacologia
11.
Nat Commun ; 10(1): 4155, 2019 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-31519912

RESUMO

Zika virus (ZIKV) infection results in an increased risk of spontaneous abortion and poor intrauterine growth although the underlying mechanisms remain undetermined. Little is known about the impact of ZIKV infection during the earliest stages of pregnancy, at pre- and peri-implantation, because most current ZIKV pregnancy studies have focused on post-implantation stages. Here, we demonstrate that trophectoderm cells of pre-implantation human and mouse embryos can be infected with ZIKV, and propagate virus causing neural progenitor cell death. These findings are corroborated by the dose-dependent nature of ZIKV susceptibility of hESC-derived trophectoderm cells. Single blastocyst RNA-seq reveals key transcriptional changes upon ZIKV infection, including nervous system development, prior to commitment to the neural lineage. The pregnancy rate of mice is >50% lower in pre-implantation infection than infection at E4.5, demonstrating that pre-implantation ZIKV infection leads to miscarriage. Cumulatively, these data elucidate a previously unappreciated association of pre- and peri-implantation ZIKV infection and microcephaly.


Assuntos
Complicações Infecciosas na Gravidez/metabolismo , Infecção por Zika virus/complicações , Infecção por Zika virus/metabolismo , Zika virus/patogenicidade , Aborto Espontâneo/metabolismo , Aborto Espontâneo/fisiopatologia , Animais , Blastocisto/citologia , Blastocisto/metabolismo , Implantação do Embrião/fisiologia , Feminino , Desenvolvimento Fetal/genética , Desenvolvimento Fetal/fisiologia , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Gravidez , RNA Viral/genética , Pesquisa Médica Translacional/métodos , Trofoblastos/citologia , Trofoblastos/metabolismo
12.
Int J Mol Sci ; 20(18)2019 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-31540219

RESUMO

During the peri-implantation period, multinucleated syncytia are formed in the sheep placenta. For over 20 years the scientific consensus has been that during trophoblast syncytialization in sheep, binucleate trophoblast giant cells (BNCs) differentiate from mononuclear trophoblast cells, and individual BNCs fuse with individual luminal epithelial (LE) cells to form trinucleate cells. These trophoblast-LE syncytial plaques then grow through continued BNC migration and fusion. Therefore, LE cells are thought to be incorporated into syncytial plaques. However, these ideas were based on electron microscopy studies, without benefit of molecular markers for BNC and LE cells to support conclusions. The aim of this study was to observe interactions between BNCs and uterine LE cells using immunohistochemical localization for molecular markers for BNCs and uterine LE cells. We performed immunofluorescence staining, laser capture microdissection, and TUNEL staining on the uterine-placental tissues of sheep during early placentation. We observed: (1) syncytial cells containing more than two nuclei within the trophoblast cell layer; (2) depolarized LE cells that express caspase 3 and stain positively for TUNEL; (3) engulfment of caspase 3-positive LE cells by trophoblast giant cells (TGCs) and empty spaces within the LE layer at sites of implantation; (4) rapid enlargement of syncytial plaques; and (5) E-cadherin and TUNEL-positive cells within the uterine stroma underlying degenerating LE was coincident with accumulation of CD45-positive cells at these sites. These data suggest that during early placentation: (1) fusion between trophoblasts is not limited to the formation of BNCs, and the term 'trophoblast giant cell (TGC)' may be appropriate; (2) LE cells undergo apoptosis; (3) apoptotic LE cells are eliminated by TGCs; (4) fusion is not limited to the incorporation of new BNCs but involves the lateral fusion between growing syncytial plaques; and (5) TGCs carry apoptotic LE cells away from the uterine-placental interface for elimination by immune cells within the stroma. These data indicate that uterine LE cells are not incorporated into syncytial plaques, but are engulfed and eliminated, and that early placentation in sheep is more similar to early placentation in humans than is currently understood in that both develop mononucleated cytotrophoblast and multinucleated syncytiotrophoblast layers of entirely placental origin. The elimination of LE cells by sheep TGCs might provide insights into elimination and penetration of LE cells during human embryo implantation.


Assuntos
Biomarcadores/metabolismo , Células Epiteliais/citologia , Células Gigantes/citologia , Placentação , Trofoblastos/citologia , Animais , Caderinas/metabolismo , Caspase 3/metabolismo , Diferenciação Celular , Fusão Celular , Movimento Celular , Células Epiteliais/metabolismo , Feminino , Células Gigantes/metabolismo , Imuno-Histoquímica , Queratinas/metabolismo , Antígenos Comuns de Leucócito/metabolismo , Placenta/citologia , Placenta/metabolismo , Gravidez , Ovinos , Trofoblastos/metabolismo
13.
Int J Mol Sci ; 20(18)2019 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-31500240

RESUMO

Omega-3 fatty acids are important to pregnancy and neonatal development and health. One mechanism by which omega-3 fatty acids exert their protective effects is through serving as substrates for the generation of specialized pro-resolving lipid mediators (SPM) that potently limit and resolve inflammatory processes. We recently identified that SPM levels are increased in maternal blood at delivery as compared to umbilical cord blood, suggesting the placenta as a potential site of action for maternal SPM. To explore this hypothesis, we obtained human placental samples and stained for the SPM resolvin D2 (RvD2) receptor GPR18 via immunohistochemistry. In so doing, we identified GPR18 expression in placental vascular smooth muscle and extravillous trophoblasts of the placental tissues. Using in vitro culturing, we confirmed expression of GPR18 in these cell types and further identified that stimulation with RvD2 led to significantly altered responsiveness (cytoskeletal changes and pro-inflammatory cytokine production) to lipopolysaccharide inflammatory stimulation in human umbilical artery smooth muscle cells and placental trophoblasts. Taken together, these findings establish a role for SPM actions in human placental tissue.


Assuntos
Ácidos Docosa-Hexaenoicos/farmacologia , Músculo Liso Vascular/citologia , Receptores Acoplados a Proteínas-G/genética , Trofoblastos/citologia , Adulto , Células Cultivadas , Ácidos Graxos Ômega-3/farmacologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , Idade Materna , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , Placenta/citologia , Placenta/efeitos dos fármacos , Placenta/metabolismo , Gravidez , Receptores Acoplados a Proteínas-G/metabolismo , Trofoblastos/efeitos dos fármacos , Trofoblastos/metabolismo , Adulto Jovem
14.
Nat Commun ; 10(1): 3557, 2019 08 07.
Artigo em Inglês | MEDLINE | ID: mdl-31391456

RESUMO

Mammalian embryos change shape dramatically upon implantation. The cellular and molecular mechanism underlying this transition are largely unknown. Here, we show that this transition is directed by cross talk between the embryonic epiblast and the first extra-embryonic tissue, the trophectoderm. Specifically, we show via visualisation of a Cdx2-GFP reporter line and pharmacologically mediated loss and gain of function experiments that the epiblast provides FGF signal that results in differential fate acquisition in the multipotent trophectoderm leading to the formation of a tissue boundary within this tissue. The trophectoderm boundary becomes essential for expansion of the tissue into a multi-layered epithelium. Folding of this multi-layered trophectoderm induces spreading of the second extra-embryonic tissue, the primitive endoderm. Together, these events remodel the pre-implantation embryo into its post-implantation cylindrical shape. Our findings uncover how communication between embryonic and extra-embryonic tissues provides positional cues to drive shape changes in mammalian development during implantation.


Assuntos
Implantação do Embrião/fisiologia , Embrião de Mamíferos/embriologia , Camadas Germinativas/embriologia , Morfogênese/fisiologia , Trofoblastos/fisiologia , Animais , Embrião de Mamíferos/diagnóstico por imagem , Feminino , Fatores de Crescimento de Fibroblastos/metabolismo , Camadas Germinativas/diagnóstico por imagem , Camadas Germinativas/metabolismo , Ligantes , Masculino , Camundongos , Camundongos Transgênicos , Microscopia de Fluorescência , Trofoblastos/metabolismo
15.
Mol Med Rep ; 20(4): 3256-3264, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31432141

RESUMO

Preeclampsia (PE) is a serious pregnancy­specific pathologic complication, and represents a primary cause of mother and fetus mortality. Abnormally expressed microRNAs (miRNAs) serve important regulatory roles in the development of PE. At present, the pathogenesis and molecular mechanism of PE remain unclear. The aim of the present study was to investigate the potential functions of miRNA (miR)­320a in the human extravillous trophoblast cell line HTR­8/SVneo and to identify the molecular mechanisms underlying miR­320a function. Reverse transcription­quantitative PCR was used in the present study to detect the levels of miR­320a in the placentas of 57 pregnant patients with PE and 57 healthy pregnant patients. The effects of miR­320a overexpression on the proliferation and invasion of HTR­8/SVneo cells were determined using MTT and Transwell invasion assays. Western blot analysis and dual luciferase reporter assay were used to identify the genes targeted by miR­320a. The present results suggested that miR­320a expression level was decreased in placentas of patients with PE and the expression level of miR­320a was found to be associated with the pathogenesis of PE (P<0.05). Overexpression of miR­320a using miR­320a mimics significantly inhibited cell proliferation and invasion in HTR­8/SVneo cells in vitro (P<0.05). Furthermore, interleukin (IL)­4 was identified to be a direct target gene of miR­320a. miR­320a could repress IL­4 expression by binding to its 3' untranslated region (P<0.05). Mechanistic studies suggested that IL­4 was a functional target gene of miR­320a, and miR­320a upregulation inhibited the proliferation and invasion of HTR­8/SVneo cells by directly targeting IL­4 (P<0.05). Collectively, to the best of our knowledge, the present study is the first to suggest that miR­320a may be a downregulated miRNA during PE, and IL­4 may act as a functional target gene of miR­320a. The present study suggested that miR­320a upregulation was involved in the development of PE by inhibiting the proliferation and invasion of trophoblast cells by targeting IL­4, indicating that the miR­320a/IL­4 pathway may represent a novel therapeutic target for PE treatment.


Assuntos
Movimento Celular , Interleucina-4/biossíntese , MicroRNAs/biossíntese , Pré-Eclâmpsia/metabolismo , Trofoblastos/metabolismo , Regulação para Cima , Adulto , Proliferação de Células , Feminino , Humanos , Pré-Eclâmpsia/patologia , Gravidez , Trofoblastos/patologia
16.
Mol Cell ; 75(6): 1103-1116.e9, 2019 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-31420216

RESUMO

The mitochondrial pathway of apoptosis is controlled by the ratio of anti- and pro-apoptotic members of the Bcl-2 family of proteins. The molecular events underlying how a given physiological stimulus changes this ratio to trigger apoptosis remains unclear. We report here that human 17-ß-estradiol (E2) and its related steroid hormones induce apoptosis by binding directly to phosphodiesterase 3A, which in turn recruits and stabilizes an otherwise fast-turnover protein Schlafen 12 (SLFN12). The elevated SLFN12 binds to ribosomes to exclude the recruitment of signal recognition particles (SRPs), thereby blocking the continuous protein translation occurring on the endoplasmic reticulum of E2-treated cells. These proteins include Bcl-2 and Mcl-1, whose ensuing decrease triggers apoptosis. The SLFN12 protein and an apoptosis activation marker were co-localized in syncytiotrophoblast of human placentas, where levels of estrogen-related hormones are high, and dynamic cell turnover by apoptosis is critical for successful implantation and placenta development.


Assuntos
Apoptose/efeitos dos fármacos , Estradiol/farmacologia , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Trofoblastos/metabolismo , Adulto , Nucleotídeo Cíclico Fosfodiesterase do Tipo 3/metabolismo , Feminino , Células HeLa , Humanos , Células MCF-7 , Proteína de Sequência 1 de Leucemia de Células Mieloides/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ribossomos/metabolismo
17.
EMBO J ; 38(18): e100849, 2019 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-31424120

RESUMO

The placenta is an autonomous organ that maintains fetal growth and development. Its multinucleated syncytiotrophoblast layer, providing fetal nourishment during gestation, exhibits characteristics of cellular senescence. We show that in human placentas from pregnancies with intrauterine growth restriction, these characteristics are decreased. To elucidate the functions of pathways regulating senescence in syncytiotrophoblast, we used dynamic contrast-enhanced MRI in mice with attenuated senescence programs. This approach revealed an altered dynamics in placentas of p53-/- , Cdkn2a-/- , and Cdkn2a-/- ;p53-/- mice, accompanied by histopathological changes in placental labyrinths. Human primary syncytiotrophoblast upregulated senescence markers and molecular pathways associated with cell-cycle inhibition and senescence-associated secretory phenotype. The pathways and components of the secretory phenotype were compromised in mouse placentas with attenuated senescence and in human placentas from pregnancies with intrauterine growth restriction. We propose that molecular mediators of senescence regulate placental structure and function, through both cell-autonomous and non-autonomous mechanisms.


Assuntos
Inibidor p16 de Quinase Dependente de Ciclina/genética , Retardo do Crescimento Fetal/genética , Redes Reguladoras de Genes , Placenta/diagnóstico por imagem , Proteína Supressora de Tumor p53/genética , Animais , Senescência Celular , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica , Humanos , Imagem por Ressonância Magnética , Camundongos , Fenótipo , Placenta/metabolismo , Gravidez , Transdução de Sinais , Trofoblastos/metabolismo
18.
Dis Markers ; 2019: 4976845, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31467616

RESUMO

Background: Preeclampsia (PE) is a pregnancy-related syndrome characterized by hypertension and proteinuria after the 20th week of gestation. The long noncoding RNAs (lncRNAs) have been recently discovered for their roles in the pathogenesis of PE. This study is aimed at determining the expression of lncRNA MIR503 host gene (MIR503HG) in PE placental tissues and exploring the molecular mechanism underlying MIR503HG-mediated trophoblast cell proliferation, invasion, and migration. Methods: The expression level of MIR503HG in placental tissues, HTR-8/SVneo, and JEG3 cells was determined by quantitative real-time PCR; western blot detected the relevant protein expression levels in HTR-8/SVneo and JEG3 cells; flow cytometry determined cell apoptosis and cell cycle of HTR-8/SVneo and JEG3 cells; trophoblast cell proliferation, invasion, and migration of HTR-8/SVneo and JEG3 cells were measured by CCK-8, transwell invasion, and wound healing assays, respectively. Results: The highly expressed MIR503HG was detected in PE placental tissues compared to normal placental tissues. MIR503HG overexpression suppressed cell proliferation, invasion, and migration of HTR-8/SVneo and JEG3 cells, while knockdown of MIR503HG increased trophoblast cell proliferation, invasion, and migration. Flow cytometry results showed that MIR503HG overexpression induced apoptosis and caused cell cycle arrest at the G0/G1 phase, while MIR503HG knockdown had the opposite actions in HTR-8/SVneo and JEG3 cells. Western blot assay results showed that MIR503HG overexpression suppressed the matrix metalloproteinase-2/-9 and the snail protein expression and increased the E-cadherin expression in trophoblast cells. In addition, MIR503HG overexpression suppressed the NF-κB signaling pathway by inhibiting the phosphorylation of IκBα and the nuclear translocation of NF-κB signaling subunit p65. On the other hand, MIR503HG knockdown played an opposite role in these protein expression levels. Conclusion: Our results showed that MIR503HG inhibited the proliferation, invasion, and migration of HTR-8/SVneo and JEG3 cells, which may be related to the pathogenesis of PE.


Assuntos
Movimento Celular , Proliferação de Células , Metaloproteinases da Matriz/metabolismo , NF-kappa B/metabolismo , Pré-Eclâmpsia/metabolismo , RNA Longo não Codificante/genética , Trofoblastos/metabolismo , Linhagem Celular Tumoral , Células Cultivadas , Feminino , Humanos , Metaloproteinases da Matriz/genética , Pré-Eclâmpsia/genética , Gravidez , RNA Longo não Codificante/metabolismo , Transdução de Sinais , Fatores de Transcrição da Família Snail/metabolismo , Trofoblastos/citologia
19.
Int J Mol Sci ; 20(17)2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31443507

RESUMO

The effectors of the type IV secretion system (T4SS) of bacteria play important roles in mediating bacterial intracellular proliferation and manipulating host-related pathway responses to bacterial infection. Brucella Spp. inhibit the apoptosis of host cells to benefit their own intracellular proliferation. However, the underlying mechanisms between T4SS effectors and Brucella-inhibited apoptosis in goat trophoblast cells remain unclear. Here, based on Brucella suis vaccine strain 2, the VceC was deleted by allelic exchange. We show that ΔVceC was able to infect and proliferate to high titers in goat trophoblast cells (GTCs) and increase C/EBP-homologous protein (CHOP)-mediated apoptosis. GRP78 expression decreased upon ΔVceC infection. In addition, we discovered that the inositolrequiring enzyme 1 (IRE1) pathway was inhibited in this process. Changing endoplasmic reticulum (ER) stress affected Brucella intracellular replication in GTCs. The replication of ΔVceC was more sensitive under the different ERstress conditions in the GTC line after treatment with ER stress inhibitors 4 phenyl butyric acid (4-PBA) or ER stress activator Tm. Together, our findings show that VceC has a protective effect on the intracellular persistence of Brucella infection, and inhibits ER stress-induced apoptosis in the CHOP pathway. The present work provides new insights for understanding the mechanism of VceC in the establishment of chronic Brucella infection.


Assuntos
Proteínas da Membrana Bacteriana Externa/metabolismo , Brucella/fisiologia , Brucelose/veterinária , Proteínas Serina-Treonina Quinases/metabolismo , Trofoblastos/metabolismo , Trofoblastos/microbiologia , Sequência de Aminoácidos , Animais , Apoptose , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/genética , Estresse do Retículo Endoplasmático/genética , Cabras , Interações Hospedeiro-Patógeno , Humanos , Viabilidade Microbiana , Mutação , Proteínas Serina-Treonina Quinases/química , Proteínas Serina-Treonina Quinases/genética , Ovinos , Doenças dos Ovinos/metabolismo , Doenças dos Ovinos/microbiologia , Transdução de Sinais
20.
Mol Cell ; 75(3): 523-537.e10, 2019 08 08.
Artigo em Inglês | MEDLINE | ID: mdl-31256989

RESUMO

Long noncoding RNAs (lncRNAs) cause Polycomb repressive complexes (PRCs) to spread over broad regions of the mammalian genome. We report that in mouse trophoblast stem cells, the Airn and Kcnq1ot1 lncRNAs induce PRC-dependent chromatin modifications over multi-megabase domains. Throughout the Airn-targeted domain, the extent of PRC-dependent modification correlated with intra-nuclear distance to the Airn locus, preexisting genome architecture, and the abundance of Airn itself. Specific CpG islands (CGIs) displayed characteristics indicating that they nucleate the spread of PRCs upon exposure to Airn. Chromatin environments surrounding Xist, Airn, and Kcnq1ot1 suggest common mechanisms of PRC engagement and spreading. Our data indicate that lncRNA potency can be tightly linked to lncRNA abundance and that within lncRNA-targeted domains, PRCs are recruited to CGIs via lncRNA-independent mechanisms. We propose that CGIs that autonomously recruit PRCs interact with lncRNAs and their associated proteins through three-dimensional space to nucleate the spread of PRCs in lncRNA-targeted domains.


Assuntos
RNA Longo não Codificante/genética , Animais , Cromatina/genética , Montagem e Desmontagem da Cromatina , Ilhas de CpG/genética , Genoma/genética , Impressão Genômica/genética , Humanos , Camundongos , Complexo Repressor Polycomb 1/genética , Regiões Promotoras Genéticas , Células-Tronco/metabolismo , Trofoblastos/metabolismo
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