RESUMO
During the process of decidualization, the stromal cells of the endometrium change dynamically to create a favorable environment for embryo implantation. Lysosome activity has often been associated with physiological changes in the endometrium during the preimplantation period and early pregnancy. In this study, the effect of para-nonylphenol (p-NP), an endocrine disruptor, on human immortalized endometrial stromal cells (tHESCs) was investigated. After exposure to p-NP (1 nM and 1 pM), the cells were examined for the decidualization markers connexin-43, insulin like growth factor binding protein 1 (IGFBP1), and prolactin. In addition, the effect of p-NP on lysosome biogenesis and exocytosis was investigated by examining the expression and localization of the transcription factor EB (TFEB) and that of the lysosomal-associated membrane protein 1 (LAMP-1). Finally, we evaluated the effect of p-NP on extracellular matrix (ECM) remodeling using a fibronectin assay. Our results showed that p-NP reduced the expression of prolactin protein, increased the nuclear localization of TFEB, and induced the increase and translocation of the lysosomal protein LAMP-1 to the membrane of tHESCs. The data indicate an impairment of decidualization and suggest an increase in lysosomal biogenesis and exocytosis, which is supported by the higher release of active cathepsin D by tHESCs. Given the importance of cathepsins in the processing and degradation of the ECM during trophoblast invasiveness and migration into the decidua, our results appear to be clear evidence of the negative effects of p-NP on endometrial processes that are fundamental to reproductive success and the establishment of pregnancy.NEW & NOTEWORTHY Endocrine disruptors, such as para-nonylphenol, affect the decidualization of human endometrial stromal cells with an impact on decidualization itself, lysosome biogenesis and exocytosis, and extracellular matrix remodeling. All these alterations may negatively impact embryo implantation with the success of reproduction and the establishment of pregnancy.
Assuntos
Endométrio , Lisossomos , Fenóis , Prolactina , Células Estromais , Humanos , Feminino , Lisossomos/metabolismo , Lisossomos/efeitos dos fármacos , Células Estromais/metabolismo , Células Estromais/efeitos dos fármacos , Fenóis/farmacologia , Fenóis/toxicidade , Endométrio/metabolismo , Endométrio/efeitos dos fármacos , Endométrio/citologia , Prolactina/metabolismo , Decídua/metabolismo , Decídua/efeitos dos fármacos , Decídua/citologia , Exocitose/efeitos dos fármacos , Implantação do Embrião/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Proteína 1 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Proteína 1 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Matriz Extracelular/metabolismo , Matriz Extracelular/efeitos dos fármacos , Gravidez , Proteína 1 de Membrana Associada ao LisossomoRESUMO
Proinflammatory cytokines are produced in pregnancy in response to the invading pathogens and/or nonmicrobial causes such as damage-associated molecules and embryonic semi-allogenic antigens. While inflammation is essential for a successful pregnancy, an excessive inflammatory response is implicated in several pathologies including pre-eclampsia (PE). This review focuses on the proinflammatory cytokine macrophage migration inhibitory factor (MIF), a critical regulator of the innate immune response and a major player of processes allowing normal placental development. PE is a severe pregnancy-related syndrome characterized by exaggerated inflammatory response and generalized endothelial damage. In some cases, usually of early onset, it originates from a maldevelopment of the placenta, and is associated with intrauterine growth restriction (IUGR) (placental PE). In other cases, usually of late onset, pre-pregnancy maternal diseases represent risk factors for the development of the disease (maternal PE). Available data suggest that low MIF production in early pregnancy could contribute to the abnormal placentation. The resulting placental hypoxia in later pregnancy could produce high release of MIF in maternal serum typical of placental PE. More studies are needed to understand the role of MIF, if any, in maternal PE.
Assuntos
Retardo do Crescimento Fetal/sangue , Oxirredutases Intramoleculares/sangue , Fatores Inibidores da Migração de Macrófagos/sangue , Placenta/metabolismo , Pré-Eclâmpsia/sangue , Feminino , Retardo do Crescimento Fetal/patologia , Humanos , Placenta/patologia , Pré-Eclâmpsia/patologia , GravidezRESUMO
Bisphenol A (BPA) is a synthetic phenol extensively used in the manufacture of polycarbonate plastics and epoxy resins and a component of liquid and food storages. Among health disorders potentially attributed to BPA, the effects on metabolism have been especially studied. BPA represents a hazard in prenatal life because of its presence in tissues and fluids during pregnancy. Our recent study in rats fed with BPA showed a placental increase in glucose type 1 transporter (GLUT-1), suggesting a higher uptake of glucose. However, the role of BPA on GLUT transporters in pregnant women with metabolic dysfunction has not yet been investigated. In this study, placental tissue from 26 overweight (OW) women and 32 age-matched normal weight (NW) pregnant women were examined for expression of GLUT1 and GLUT4. Placental explants from OW and NW mothers were exposed to BPA 1 nM and 1 µM and tested for GLUTs expression. The data showed a different response of placental explants to BPA in GLUT1 expression with an increase in NW mothers and a decrease in OW ones. GLUT4 expression was lower in the explants from OW than NW mothers, while no difference was showed between OW and NW in placental biopsies for any of the transporters.
Assuntos
Compostos Benzidrílicos/toxicidade , Transportador de Glucose Tipo 1/metabolismo , Transportador de Glucose Tipo 4/metabolismo , Sobrepeso/complicações , Fenóis/toxicidade , Placenta/efeitos dos fármacos , Complicações na Gravidez/induzido quimicamente , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Sobrepeso/metabolismo , Placenta/metabolismo , Gravidez , Complicações na Gravidez/metabolismoRESUMO
BACKGROUND: Toxoplasma gondii is a protozoan parasite that causes congenital toxoplasmosis by transplacental transmission. Parasite strains are genetically diverse and disease severity is related to the genotype. In Uberlândia city, Brazil, two virulent strains were isolated: TgChBrUD1 and TgChBrUD2. Congenital toxoplasmosis is more prevalent in South America compared to Europe, and more often associated with severe symptoms, usually as a result of infection with atypical strains. METHODS: Considering that T. gondii has shown high genetic diversity in Brazil, the effectiveness of traditional treatment may not be the same, as more virulent strains of atypical genotypes may predominate. Thus, the aim of this study were to evaluate the Brazilian strain infection rate in human villous explants and the azithromycin efficacy with regard to the control of these strains compared to traditional therapy. Villi were infected with RH, ME49, TgChBrUD1 or TgChBrUD2 strains and treated with azithromycin, spiramycin or a combination of pyrimethamine plus sulfadiazine. The villous viability was analyzed by LDH assay and morphological analysis. Parasite proliferation, as well as production of cytokines was analyzed by qPCR and ELISA, respectively. Statistical analysis was performed using the GraphPad Prism 5.0. RESULTS: The treatments were not toxic and TgChBrUD1 infected villi showed a higher parasite burden compared with others strains. Treatments significantly reduced the intracellular proliferation of T. gondii, regardless of the strain. TgChBrUD1-infected villi produced a larger amount of MIF, IL-6 and TGF-ß1 compared with other infected villi. Azithromycin treatment increased MIF production by RH- or TgChBrUD2-infected villi, but in ME49- or TgChBrUD1-infected villi, the MIF production was not altered by treatment. On the other hand, azithromycin treatment induced lower IL-6 production by ME49- or TgChBrUD1-infected villi. CONCLUSIONS: Azithromycin treatment was effective against T. gondii Brazilian strains compared with conventional treatment. Also, the TgChBrUD1 strain replicated more in villi and modulated important cytokines involved in parasite control, showing that different strains use different strategies to evade the host immune response and ensure their survival.
Assuntos
Azitromicina/farmacologia , Coccidiostáticos/farmacologia , Citocinas/metabolismo , Placenta/parasitologia , Toxoplasma/efeitos dos fármacos , Brasil , Feminino , Humanos , Gravidez , Terceiro Trimestre da GravidezRESUMO
This study was conducted to investigate annexin A1 (ANXA1) functions in human placental explants infected with Toxoplasma gondii (T. gondii). We examined the first and third trimester placental explants infected with T. gondii (nâ¯=â¯7 placentas/group) to identify the number and location of parasites, ANXA1 protein, potential involvement of formyl peptide receptors (FPR1 and FPR2), and COX-2 expressions by immunohistochemistry. Treatments with Ac2-26 mimetic peptide of ANXA1 were performed to verify the parasitism rate (ß-galactosidase assay), prostaglandin E2 levels (ELISA assay), and ANXA1, FPR1 and COX-2 expression in third trimester placentas. Placental explants of third trimester expressed less ANXA1 and were more permissive to T. gondii infection than first trimester placentas that expressed more ANXA1. Ac2-26 treatment increases endogenous ANXA1 and decreases parasitism rate, COX-2, and prostaglandin E2 levels. Altogether, these data provide further insight into the anti-parasitic and anti-inflammatory effects of ANXA1 in placentas infected with T. gondii.
Assuntos
Anexina A1/farmacologia , Antiparasitários/farmacologia , Placenta/efeitos dos fármacos , Toxoplasma/efeitos dos fármacos , Toxoplasma/patogenicidade , Toxoplasmose/tratamento farmacológico , Anti-Inflamatórios/farmacologia , Estudos Transversais , Ciclo-Oxigenase 2/metabolismo , Dinoprostona/metabolismo , Feminino , Humanos , Inflamação/tratamento farmacológico , Peptídeos/farmacologia , Placenta/patologia , Placenta/fisiopatologia , Gravidez , Terceiro Trimestre da Gravidez , Receptores de Formil Peptídeo/metabolismo , Receptores de Lipoxinas/metabolismo , Toxoplasmose/patologia , beta-Galactosidase/análiseRESUMO
Human chorionic gonadotropin (hCG) is a hormone of considerable importance in the establishment, promotion and maintenance of human pregnancy. It has been clearly demonstrated that hCG exerts multiple endocrine, paracrine and autocrine actions on a variety of gestational and non-gestational cells and tissues. These actions are directed to promote trophoblast invasiveness and differentiation, placental growth, angiogenesis in uterine vasculature, hormone production, modulation of the immune system at the maternal-fetal interface, inhibition of myometrial contractility as well as fetal growth and differentiation. In recent years, considerable interest has been raised towards the biological effects of environmental contaminants, particularly endocrine disrupting chemicals (EDCs). Emerging evidence suggests that prenatal exposure to selected EDCs can have a deleterious impact on the fetus and long-lasting consequences also in adult life. The results of the in vitro effects of commonly found EDCs, particularly Bisphenol A (BPA) and para-Nonylphenol (p-NP), indicate that these substances can alter hCG production and through this action could exert their fetal damage, suggesting that hCG could represent and become a potentially useful clinical biomarker of an inappropriate prenatal exposure to these substances.
Assuntos
Gonadotropina Coriônica/metabolismo , Disruptores Endócrinos/toxicidade , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Feminino , Humanos , Exposição Materna/efeitos adversos , Gravidez , Efeitos Tardios da Exposição Pré-Natal/etiologia , Efeitos Tardios da Exposição Pré-Natal/patologia , Trofoblastos/efeitos dos fármacos , Trofoblastos/metabolismoRESUMO
Trophoblast infection by Toxoplasma gondii plays a pivotal role in the vertical transmission of toxoplasmosis. Here, we investigate whether the antibiotic therapy with azithromycin, spiramycin and sulfadiazine/pyrimethamine are effective to control trophoblast infection by two Brazilian T. gondii genotypes, TgChBrUD1 or TgChBrUD2. Two antibiotic protocols were evaluated, as follow: i) pre-treatment of T. gondii-tachyzoites with selected antibiotics prior trophoblast infection and ii) post-treatment of infected trophoblasts. The infection index/replication and the impact of the antibiotic therapy on the cytokine milieu were characterized. It was observed that TgChBrUD2 infection induced lower infection index/replication as compared to TgChBrUD1. Regardless the therapeutic protocol, azithromycin was more effective to control the trophoblast infection with both genotypes when compared to conventional antibiotics. Azithromycin induced higher IL-12 production in TgChBrUD1-infected cells that may synergize the anti-parasitic effect. In contrast, the effectiveness of azithromycin to control the TgChBrUD2-infection was not associated with the IL-12 production. BeWo-trophoblasts display distinct susceptibility to T. gondii genotypes and the azithromycin treatment showed to be more effective than conventional antibiotics to control the T. gondii infection/replication regardless the parasite genotype.
Assuntos
Antiprotozoários/farmacologia , Azitromicina/farmacologia , Toxoplasma/efeitos dos fármacos , Trofoblastos/parasitologia , Linhagem Celular Tumoral , Citocinas/metabolismo , Combinação de Medicamentos , Genótipo , Humanos , Interleucina-12/metabolismo , Pirimetamina/farmacologia , Espiramicina/farmacologia , Sulfadiazina/farmacologia , Toxoplasma/classificação , Toxoplasma/genética , Toxoplasma/imunologia , Trofoblastos/efeitos dos fármacosRESUMO
Considering that interleukin 10 (IL10), transforming growth factor beta1 (TGFB1), and interferon gamma (IFNG) are involved in the susceptibility of BeWo trophoblast cells to Toxoplasma gondii infection, the aim of the present study was to investigate the effector mechanisms triggered by these cytokines in the control of T. gondii in BeWo cells. For this purpose, infected/uninfected BeWo cells were treated with IL10, TGFB1 (50 ng/ml), and IFNG (20 or 100 ng/ml) in order to verify the phosphorylation of signal transducers and activators of transcription 1 (STAT1), STAT3, and Smad2, parasite intracellular proliferation, as well as the Th1/Th2/IL17A cytokine production. The treatment of BeWo cells with IL10 and TGFB1 favored T. gondii proliferation, and these findings were associated with STAT3 and Smad2 phosphorylation, respectively (P < 0.05). Also, these cytokine treatments were able to down-modulate TNF alpha (TNFA) and IL6 production (P < 0.05). Low concentration of IFNG was unable to control T. gondii infection but was able to trigger STAT1 phosphorylation and up-regulate IL6 and IL17A production; whereas a high concentration of IFNG was unable to activate STAT1 but down-modulated IL6 and TNFA and increased T. gondii proliferation (P < 0.05). IL10, TGFB1, and IFNG regulate a differential T. gondii proliferation in BeWo cells because they distinctly trigger intracellular signaling pathways and cytokine production, especially IL6 and TNFA. Our data open new windows to understand the mechanisms triggered by IL10, TGFB1, and IFNG at the maternal-fetal interface in the presence of T. gondii, contributing to recognizing the importance of these effector mechanisms involved in the vertical transmission of this parasite.
Assuntos
Citocinas/metabolismo , Interferon gama/farmacologia , Interleucina-10/farmacologia , Transdução de Sinais/efeitos dos fármacos , Toxoplasmose/prevenção & controle , Fator de Crescimento Transformador beta1/farmacologia , Trofoblastos/efeitos dos fármacos , Trofoblastos/parasitologia , Linhagem Celular Tumoral , Coriocarcinoma/patologia , Suscetibilidade a Doenças , Feminino , Humanos , Técnicas In Vitro , Interleucina-16/metabolismo , Fosforilação , Gravidez , Fator de Transcrição STAT1/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/fisiologia , Proteína Smad2/metabolismo , Toxoplasma/isolamento & purificação , Toxoplasmose/metabolismo , Toxoplasmose/patologia , Trofoblastos/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Neoplasias Uterinas/patologiaRESUMO
The human endometrium is a fertility-determining tissue and a target of steroid hormones' action. Endocrine disruptors (EDs) can exert adverse effects on the physiological function of the decidua at the maternal-fetal interface. We examined the potential effects of an ED, bisphenol A (BPA), on endometrial maturation/decidualization, receptivity, and secretion of decidual factors (biomarkers). In vitro decidualized, endometrial stromal cells from six hysterectomy specimens were treated with 1â pM-1 âµM of BPA, for 24 âh and assessed for cell viability and proliferation. Three non-toxic concentrations of BPA (1 âµM, 1 ânM, and 1 âpM) were selected to study its influence on secretion of cell decidualization biomarkers (IGF-binding protein and decidual prolactin (dPRL)), macrophage migration inhibitory factor (MIF) secretion, and hormone receptors' expression (estrogen receptors (ERα and ERß); progesterone receptors (PRA and PRB); and human chorionic gonadotropin (hCG)/LH receptor (LH-R)). The results showed a decrease in cell viability (P<0.001) in response to BPA at the level of 1â mM. At the non-toxic concentrations used, BPA perturbed the expression of ERα, ERß, PRA, PRB, and hCG/LH-R (P<0.05). Furthermore, 1 âµM of BPA reduced the mRNA transcription of dPRL (P<0.05). Secretion of MIF was stimulated by all BPA treatments, the lowest concentration (1 âpM) being the most effective (P<0.001). The multi-targeted disruption of BPA on decidual cells, at concentrations commonly detected in the human population, raises great concern about the possible consequences of exposure to BPA on the function of decidua and thus its potential deleterious effect on pregnancy.
Assuntos
Compostos Benzidrílicos/toxicidade , Decídua/citologia , Decídua/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Poluentes Ambientais/toxicidade , Fenóis/toxicidade , Antimetabólitos/farmacologia , Biomarcadores/análise , Biomarcadores/metabolismo , Bromodesoxiuridina/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Decídua/metabolismo , Endométrio/citologia , Endométrio/efeitos dos fármacos , Endométrio/crescimento & desenvolvimento , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Técnicas In Vitro , Proteína 1 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Fatores Inibidores da Migração de Macrófagos/metabolismo , Ciclo Menstrual/efeitos dos fármacos , Gravidez , RNA Mensageiro/biossíntese , RNA Mensageiro/genéticaRESUMO
OBJECTIVES: To investigate serum levels, tissue/cellular expression of macrophage migration inhibitory factor (MIF) in patients with limited (lSSc) and diffuse (dSSc) systemic sclerosis. METHODS: 10 lSSc-patients, 10 dSSc-patients and 10 controls were enrolled. MIF serum levels were assayed by ELISA. MIF and its receptors CD74/CD44 were evaluated by immunohistochemistry on skin biopsies from patients with dSSc, lSSc (affected and not-affected skin) and controls. MIF levels were assessed (ELISA) in supernatants of healthy dermal microvascular endothelial cells (MVECs) and in control (CTR), non-affected SSc (NA) and affected (SSc) fibroblasts treated for 48 h with 10% control serum and 10% SSc-serum. MIF supernatant (ELISA) and mRNA (quantitative real-time PCR) levels were determined in SSc dermal fibroblasts and in control dermal fibroblasts untreated or stimulated at 6 h-24 h-48 h with bleomycin (50 mU/ml). RESULTS: Serum MIF was significantly higher in dSSc (18.7±4.1 ng/ml, p<0.001) and in lSSc (10.4±4.4 ng/ml, p<0.001) patients respect to controls (2.6±1.4 ng/ml). Enhanced MIF immunoreactivity was found in keratinocytes, fibroblasts, endothelium, sebaceous/sweat glands from lSSc/dSSc affected skin. Faint MIF immunoreactivity was found in control skin and not-affected skin of lSSc patients. No differences were found in CD74/CD44 receptors' analysis among control and dSSc/lSSc affected and non-affected skin. MVECs and fibroblasts (CTR, NA and SSc) produced significantly more MIF, when stimulated with SSc serum respect to control-serum (p<0.001). Finally, MIF mRNA levels significantly increased at 6h (p<0.001) and decreased at 48 h (p<0.001) in control fibroblasts treated with bleomycin compared to control untreated. Simultaneously, MIF supernatant protein levels increased after 48 h (p<0.01) in bleomycin-treated fibroblasts respect to untreated ones. CONCLUSIONS: These results suggest that MIF could be implicated in the pathogenesis of SSc, probably acting as protective factor against the SSc stressful conditions.
Assuntos
Fibroblastos/metabolismo , Oxirredutases Intramoleculares/sangue , Fatores Inibidores da Migração de Macrófagos/sangue , Esclerodermia Difusa/sangue , Esclerodermia Limitada/sangue , Pele/metabolismo , Adulto , Antígenos de Diferenciação de Linfócitos B/metabolismo , Biomarcadores/sangue , Biópsia , Bleomicina/farmacologia , Estudos de Casos e Controles , Células Cultivadas , Células Endoteliais/imunologia , Células Endoteliais/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Fibroblastos/efeitos dos fármacos , Fibroblastos/imunologia , Antígenos de Histocompatibilidade Classe II/metabolismo , Humanos , Receptores de Hialuronatos/metabolismo , Imuno-Histoquímica , Oxirredutases Intramoleculares/genética , Fatores Inibidores da Migração de Macrófagos/genética , RNA Mensageiro/metabolismo , Esclerodermia Difusa/diagnóstico , Esclerodermia Difusa/genética , Esclerodermia Difusa/imunologia , Esclerodermia Limitada/diagnóstico , Esclerodermia Limitada/genética , Esclerodermia Limitada/imunologia , Pele/efeitos dos fármacos , Pele/imunologia , Fatores de TempoRESUMO
Gout and hyperuricemia are characterized by high uric acid levels, and their treatment involves medications that have adverse effects. In this study, we evaluated oral liposomal formulations with eremantholide C and goyazensolide as a novel approach to reduce the toxicity associated with these substances while maintaining their anti-hyperuricemic activity. We characterized the formulations and evaluated them based on encapsulation efficiency and stability over 12 months and under simulated physiological environments. We determined the toxicity of the liposomal formulations in Caco-2 cells and the anti-hyperuricemic activity in rats. The formulations exhibited nanometric size, a narrow size distribution, and a negative zeta potential, indicating their stability and uniformity. The efficient encapsulation of the sesquiterpene lactones within the liposomes emphasizes their potential for sustained release and therapeutic efficacy. Stability evaluation revealed a small decrease in the eremantholide C concentration and a remarkable stability in the goyazensolide concentration. In Caco-2 cells, the liposomes did not exert toxicity, but did exhibit an antiproliferative effect. In vivo assays demonstrated that the liposomes reduced serum uric acid levels. Our study represents an advancement in gout and hyperuricemia treatment. The liposomal formulations effectively reduced the toxicity associated with the sesquiterpene lactones while maintaining their therapeutic effects.
Assuntos
Artrite Gotosa , Hidrocarbonetos Aromáticos com Pontes , Furanos , Gota , Hiperuricemia , Sesquiterpenos , Sesterterpenos , Humanos , Ratos , Animais , Lipossomos/uso terapêutico , Ácido Úrico/uso terapêutico , Hiperuricemia/tratamento farmacológico , Células CACO-2 , Gota/tratamento farmacológico , Lactonas/farmacologia , Lactonas/uso terapêuticoRESUMO
It is becoming increasingly apparent that the dynamics of glycans reflect the physiological state of cells involved in several cell functions including growth, response to signal molecules, migration, as well as adhesion to, interaction with, and recognition of other cells. The presence of glycoconjugates in human placenta suggests their major role in maternal-fetal exchanges, intercellular adhesion, cellular metabolism, and villous vessel branching. Although several studies have described glycoconjugate distribution in the human placenta descriptions of their physiological function and control mechanisms during placental development are lacking. In this study we investigated the developmental distribution and regulation of placental core 1 O- and N-glycans focusing on early and late first trimester human pregnancy. To define the control mechanisms of the oligosaccharide chains during early placentation process, chorionic villous explants and human trophoblast cell lines were exposed to various oxygen levels. We found that oxygen tension regulates changes in core-1 O-glycan (the disaccharide Galß1-3GalNAc) epitope expression levels. Moreover, by double affinity chromatography and subsequent analysis with mass spectrometry, we identified in the heat shock protein 90-α (HSP90α) a good candidate as carrier of the Galß1-3GalNAc epitope at low oxygen tension. Our results support a fundamental role of oxygen tension in modulating glycosylation of proteins during placental development.
Assuntos
Epitopos/metabolismo , Proteínas de Choque Térmico HSP90/metabolismo , Oxigênio/fisiologia , Placenta/metabolismo , Sialiltransferases/metabolismo , Linhagem Celular Transformada , Feminino , Humanos , Técnicas de Cultura de Órgãos , Oxigênio/farmacologia , Placenta/efeitos dos fármacos , Gravidez , beta-Galactosídeo alfa-2,3-SialiltransferaseRESUMO
Embryo implantation is a complex and highly coordinated process that involves an intricate network of factors establishing intimate contact at the maternal-fetal interface. Knowledge of the human implantation process is compromised by both ethical issues, which do not allow the study of this process in vivo, and by the accuracy and reproducibility of in vitro models of human endometrium. Effective and reliable embryo implantation models are, therefore, necessary to mimic the molecular event cascade that occurs in vivo. 3D models are considered a new step to foster precision medicine and an advanced tool for the study of endometrial biology, endometrium associated diseases and to understand the complex mechanisms surrounding endometrium-embryo crosstalk. In this review we explore the various methods by which 3D cultures of endometrium and trophoblast can be created, exploring targets and applications of these in vitro models.
Assuntos
Implantação do Embrião , Trofoblastos , Feminino , Humanos , Reprodutibilidade dos Testes , EndométrioRESUMO
Congenital toxoplasmosis, caused by the opportunistic protozoan parasite T. gondii, can cause stillbirths, miscarriages and fetal abnormalities, as well as encephalitis and chorioretinitis in newborns. Available treatment options rely on antiparasitic drugs that have been linked to serious side effects, high toxicity and the development of drug-resistant parasites. The search for alternative therapeutics to treat this disease without acute toxicity for the mother and child is essential for the advancement of current therapeutic procedures. The present study aimed to unravel the mode of the anti-T. gondii action of Rottlerin, a natural polyphenol with multiple pharmacological properties described. Herein, we further assessed the antiparasitic activity of Rottlerin against T. gondii infection on the human trophoblastic cells (BeWo cells) and, for the first time, on human villous explants. We found that non-cytotoxic doses of Rottlerin impaired early and late steps of parasite infection with an irreversible manner in BeWo cells. Rottlerin caused parasite cell cycle arrest in G1 phase and compromised the ability of tachyzoites to infect new cells, thus highlighting the possible direct action on parasites. An additional and non-exclusive mechanism of action of Rottlerin involves the modulation of host cell components, by affecting lipid droplet formation, mitochondrial function and upregulation of the IL-6 and MIF levels in BeWo cells. Supporting our findings, Rottlerin also controlled T. gondii proliferation in villous explants with low toxicity and reduced the IL-10 levels, a cytokine associated with parasite susceptibility. Collectively, our results highlighted the potential use of Rottlerin as a promising tool to prevent and/or treat congenital toxoplasmosis.
RESUMO
The proinflammatory cytokine MIF (macrophage migration inhibitory factor) is involved in physiological and pathological processes in pregnancy. MIF maternal serum levels are increased in preeclampsia (PE). We hypothesize that pregnancy tissues are the source of MIF overexpression in PE. MIF protein was studied in maternal sera, placental tissues, fetal membranes, and umbilical cord of 8 control and 20 PE pregnancies: 10 with normal fetal growth (PE-AGA) and 10 with fetal growth restriction (PE-FGR). MIF levels were significantly higher in PE-AGA membranes than in controls and PE-FGR. In PE-FGR, MIF cord concentrations were higher than in PE-AGA while MIF placental levels were lower than in controls. MIF maternal serum levels were higher in PE, compared to controls, and the difference was mainly due to PE-FGR samples. These data support MIF involvement in PE pathogenesis and suggest that different pregnancy tissues contribute to MIF production in PE with and without fetoplacental compromise.
Assuntos
Membranas Extraembrionárias/metabolismo , Oxirredutases Intramoleculares/metabolismo , Fatores Inibidores da Migração de Macrófagos/metabolismo , Placenta/metabolismo , Pré-Eclâmpsia/diagnóstico , Pré-Eclâmpsia/imunologia , Adulto , Ensaio de Imunoadsorção Enzimática , Feminino , Retardo do Crescimento Fetal , Regulação da Expressão Gênica , Humanos , Imuno-Histoquímica , Oxirredutases Intramoleculares/genética , Fatores Inibidores da Migração de Macrófagos/genética , Especificidade de Órgãos , Pré-Eclâmpsia/patologia , Pré-Eclâmpsia/fisiopatologia , Gravidez , Cordão Umbilical/metabolismoRESUMO
Dietary exposure to Bisphenol A (BPA), an industrial chemical present in food containers, affects nutrient metabolism in the myocardium of offspring during intrauterine life. Using a murine model, we observed that fetal hearts from mothers exposed to BPA (2.5 µg/kg/day) for 20 days before mating and for all of the gestation had decreased expression of glucose transporter-1 (GLUT1), the principal sugar transporter in the fetal heart, and increased expression of fatty acid cluster of differentiation 36 transporter (CD36), compared to control fetuses from vehicle-treated mothers. We confirmed the suppression of GLUT1 by exposing fetal heart organotypic cultures to BPA (1 nM) for 48 h but did not detect changes in CD36 compared to controls. During pregnancy, the placenta continuously releases extracellular vesicles such as exosomes into fetal circulation. These vesicles influence the growth and development of fetal organs. When fetal heart cultures were treated with cord blood-derived exosomes isolated from BPA-fed animals, GLUT1 expression was increased by approximately 40%. Based on our results, we speculate that exosomes from cord blood, in particular placenta-derived nanovesicles, could contribute to the stabilization of the fetal heart metabolism by ameliorating the harmful effects of BPA on GLUT1 expression.
Assuntos
Compostos Benzidrílicos , Exossomos , Sangue Fetal , Transportador de Glucose Tipo 1 , Miocárdio , Fenóis , Animais , Feminino , Gravidez , Ratos , Exossomos/efeitos dos fármacos , Exossomos/metabolismo , Ácidos Graxos/metabolismo , Sangue Fetal/efeitos dos fármacos , Sangue Fetal/metabolismo , Feto/metabolismo , Transportador de Glucose Tipo 1/metabolismo , Miocárdio/metabolismo , Compostos Benzidrílicos/efeitos adversos , Fenóis/efeitos adversos , DietaRESUMO
Women with multiple sclerosis (MS) can safely become pregnant and give birth, with no side effects or impediments. Pregnancy is generally accepted as a period of well-being in which relapses have a softer evolution, particularly in the third trimester. Herein, we hypothesized that the placenta, via its "secretome", could contribute to the recognized beneficial effects of pregnancy on MS activity. We focused on a well-known receptor/ligand/decoy receptor system, such as the one composed by the receptor activator of nuclear factor-kB (RANK), its ligand (RANKL), and the decoy receptor osteoprotegerin (OPG), which have never been investigated in an integrated way in MS, pregnancy, and placenta. We reported that pregnancy at the term of gestation influences the balance between circulating RANKL and its endogenous inhibitor OPG in MS women. We demonstrated that the placenta at term is an invaluable source of homodimeric OPG. By functional studies on astrocytes, we showed that placental OPG suppresses the mRNA expression of the CCL20, a chemokine responsible for Th17 cell recruitment. We propose placental OPG as a crucial molecule for the recognized beneficial effect of late pregnancy on MS and its potential utility for the development of new and more effective therapeutic approaches.
Assuntos
Esclerose Múltipla , Feminino , Humanos , Ligantes , Esclerose Múltipla/metabolismo , Osteoprotegerina/metabolismo , Placenta/metabolismo , Gravidez , Ligação Proteica , Ligante RANK/metabolismo , Receptor Ativador de Fator Nuclear kappa-B/metabolismoRESUMO
Most studies on the mucosal immunity in female reproductive tissues have been performed in mammals. In all species, apart from their reproductive strategies, immunity in the genital mucosa is required to defend the host against luminal pathogens. In this study we investigated the role of the innate immunity of the oviductal mucosa of Xenopus laevis, an amphibian characterized by external fertilization. In particular we examined the expression and localization of Interleukin-1ß (IL1B), Macrophage migration inhibitory factor (MIF) and Interleukin-1 receptor type 1 (IL1R1) in different oviductal portions including an upper glandular region, an intermediate and a lower aglandular region (the ovisac). Tissues were examined by immunohistochemistry and western blot using polyclonal antibodies against human molecules. IL1B, MIF and IL1R1 were all shown in the three oviductal regions examined, albeit with a general increase towards the external environment. A substantial difference among the cytokine components was also observed mainly in the epithelium of the glandular and intermediate regions. Specifically, all three molecules were expressed by the luminal ciliated cells while only IL1R1 was present in the unciliated cells at the bottom of the epithelial ingrowths. The expression of IL1R1 in these cells appeared as a continuous layer separating the epithelium from the underlying tissues. While supporting the role of the innate immune system for host's defense against pathogens, the peculiar distribution of the cytokine components in the oviduct of X. laevis suggests novel immunologic strategies useful to assure gland secretion essential for egg formation and fertilization.
Assuntos
Citocinas/metabolismo , Imunidade nas Mucosas , Oviductos/imunologia , Animais , Citocinas/análise , Feminino , Imuno-Histoquímica , Interleucina-1beta/análise , Interleucina-1beta/metabolismo , Fatores Inibidores da Migração de Macrófagos/análise , Fatores Inibidores da Migração de Macrófagos/metabolismo , Oviductos/anatomia & histologia , Receptores Tipo I de Interleucina-1/análise , Receptores Tipo I de Interleucina-1/metabolismo , Xenopus laevisRESUMO
During pregnancy, both the maternal endometrium and the blastocyst have highly glycosylated proteins with glycosylations controlled in a specific manner. Carbohydrates play a fundamental role in cell-cell and cell-matrix recognition and are involved in defining the structure and integrity of tissues. The uterus' secretions, which are rich in glycoproteins and glycogen and the presence of a functional glycocalyx on the uterine epithelium, establish a favourable milieu, which is essential for the correct implantation and subsequent development of the blastocyst. Likewise, carbohydrate residues such as fucose and sialic acid present at the placental level are determinant in creating an immuno-environment, which supports the mother's tolerance towards the fetal antigens. In this review, we explore the literature concerning the role of important glycan-epitopes at the feto-maternal interface in the human species. Moreover, we also show some unpublished interesting results on changes of glycan residues in human placenta tissues from the first trimester of pregnancy.
Assuntos
Troca Materno-Fetal , Polissacarídeos/metabolismo , Endométrio/metabolismo , Feminino , Glicosilação , Humanos , Lectinas/metabolismo , Placenta/metabolismo , Polissacarídeos/química , GravidezRESUMO
OBJECTIVE: To assess the role of embryo secretome in modifying the molecular profile of glycodelin A (GdA) in endometrial organoids (ORG) mimicking the implantation window. To verify whether the use of embryo-conditioned culture medium at the time of the embryo transfer may increase in vitro fertilization outcome. DESIGN: Molecular study with human endometrial ORG and embryo-conditioned culture medium. Retrospective study using prospectively recorded data. SETTING: University hospital. PATIENT(S): For isolation and culture of endometrial glandular ORG, endometrial biopsy specimens from five white women of proven fertility undergoing laparoscopy for tubal sterilization. A total of 75 women undergoing intracytoplasmic sperm injection for tubal and/or male infertility factor. INTERVENTIONS(S): In vitro fertilization. MAIN OUTCOME MEASURE(S): Pinopodes presence in human endometrial ORG. Glycodelin A expression profile by means of two-dimensional electrophoresis. In vitro fertilization outcome. RESULT(S): This in vitro study demonstrated that the treatment of endometrial ORG with the secretome of medium conditioned by the growing embryo increased the GdA relative abundance and induced a different glycoform pattern. Biochemical and clinical pregnancy rate significantly increased when the spent medium was loaded during the transfer (17.5% vs. 36.6% and 16.5% vs. 35.1%, respectively). CONCLUSION(S): This study demonstrated that the secretome of implanting embryos is able to induce the expression as well as to determine the relative abundance and the glycosilation profile of endometrial GdA, a protein having a key role in the embryo-endometrial cross talk. Moreover, a significant increase in pregnancy rate was observed when the embryo transfer was performed by using the culture medium conditioned by the growing embryo.