RESUMO
INTRODUCTION: Macrophage migration inhibitory factor (MIF) participates in the immune response to Toxoplasma gondii, triggers ERK1/2 and prostaglandin E2 (PGE2) activation, but there is limited information on these mechanisms in human trophoblast. The present study aimed to verify the role of MIF in the ERK1/2 phosphorylation and PGE2 production, as well as its effect on the susceptibility to T. gondii in BeWo cells. METHODS: BeWo cells were treated with increasing concentrations of recombinant MIF (rMIF) and/or T. gondii-soluble tachyzoite antigen (STAg) and analyzed for ERK1/2 phosphorylation and PGE2 production by Western blotting and ELISA, respectively. Cells were also treated with increasing concentrations of rMIF, rPGE2, or ERK1/2 inhibitor and tested for T. gondii proliferation. The supernatants of cells treated with rPGE2 were assayed for cytokine production by ELISA or CBA. RESULTS: ERK1/2 phosphorylation and PGE2 production increased when the cells were treated with low MIF concentrations while the parasitism control occurred only at high MIF concentrations. STAg was unable to change ERK1/2 phosphorylation or PGE2 release. BeWo cells demonstrated increased T. gondii proliferation and reduced production of pro-inflammatory cytokines when treated with PGE2, while PD98059 diminished the parasite proliferation. DISCUSSION: The intracellular mechanisms triggered by MIF are dose-dependent in BeWo cells, and PGE2 is an important factor for the persistence of T. gondii at the maternal fetal interface. CONCLUSION: MIF was unable to control T. gondii infection in BeWo cells at low concentrations since ERK1/2 and PGE2 expression were activated, demonstrating a critical effect of these mediators favoring parasite proliferation.
Assuntos
Dinoprostona/biossíntese , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fatores Inibidores da Migração de Macrófagos/administração & dosagem , Toxoplasma/imunologia , Trofoblastos/metabolismo , Antígenos de Protozoários/farmacologia , Linhagem Celular Tumoral , Dinoprostona/farmacologia , MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , Feminino , Flavonoides/farmacologia , Humanos , Fosforilação , Toxoplasma/crescimento & desenvolvimento , Toxoplasmose/imunologia , Trofoblastos/parasitologiaRESUMO
The identification of reproductive toxicants is a major scientific challenge for human health. We investigated the effects of a selected group of environmental polluting chemicals mostly provided with estrogenic activity on the human trophoblast cell lines BeWo and HTR-8/SVneo. Cells were exposed for 24h to various concentrations (from 0.1 pM to 1 mM) of atrazine (ATR), diethylstilbestrol (DES), para-nonylphenol (p-NP), resveratrol (RES) and 17 ß-estradiol (E2) and assayed for cell viability and human beta-Chorionic Gonadotropin (ß-hCG) secretion. Decrease of cell viability as respect to control, vehicle-treated, cultures was obtained for all chemicals in the concentration range of 1 µM-1 mM in both cell types. A parallel decrease of ß-hCG secretion was observed in BeWo cells, at 1 µM-1 mM concentrations, with the only exception of ATR which caused an increase at concentrations up to 1mM. ß-hCG release was also unexpectedly inhibited by ATR, DES, p-NP and RES at non-toxic (pM-nM) concentrations. These findings raise concern about the negative, potential effects of various environmental polluting chemicals on pregnancy success and fetal health.
Assuntos
Poluentes Ambientais/toxicidade , Estradiol/toxicidade , Estrogênios/toxicidade , Trofoblastos/efeitos dos fármacos , Atrazina/toxicidade , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Gonadotropina Coriônica/metabolismo , Dietilestilbestrol/toxicidade , Humanos , Fenóis/toxicidade , Resveratrol , Estilbenos/toxicidade , Trofoblastos/metabolismoRESUMO
The female reproductive system of the tsetse fly Glossina morsitans morsitans is analysed by scanning electron microscopy (SEM). The study focuses in particular on the choriothete, a peculiar uterine structure involved in the viviparous mode of reproduction of Glossina morsitans morsitans. Under light microscopy, the choriothete appears formed by numerous tongue-like folds projecting towards the uterine lumen and lined by a thin cuticle. SEM analysis highlights for the first time a distinctive new feature that is not visible by traditional histological methods. That is a cuticular covering of the choriothete, which shows numerous thorns in the form of crest-like structures arranged in nearly parallel lines. The role of the choriothete in pregnancy and in larval nourishment is discussed.
Assuntos
Genitália Feminina/ultraestrutura , Moscas Tsé-Tsé/ultraestrutura , Viviparidade não Mamífera/fisiologia , Animais , Feminino , Genitália Feminina/citologia , Larva/citologia , Larva/ultraestrutura , Moscas Tsé-Tsé/citologiaRESUMO
ATP binding cassette transporter A1 (ABCA1) is a membrane transporter which performs cellular efflux of cholesterol and phospholipid. ABCA1's cholesterol transporting role in human placenta appears to be crucial for normal fetal development. Despite the critical importance of cholesterol in fetal development, expression of ABCA1 in the human placenta throughout gestation and its specific cellular localization have not been known yet. We therefore investigated ABCA1 expression in human placenta at first trimester and term by western blot and quantitative real-time PCR (qRT-PCR) analysis. Furthermore, its localization was investigated by immunohistochemistry and confocal microscopy. Expression of ABCA1 did not differ significantly between first trimester and term placenta at both protein and mRNA levels. Immunohistochemical data demonstrated that ABCA1 was widely localized in the villous and extravillous cytotrophoblast as well as in some stromal and endothelial cells. Confocal microscopy imaging data showed that ABCA1 was localized largely at the basolateral and to some extent at the apical side of first trimester villous cytotrophoblast cell membranes. Placental expression of ABCA1 throughout the gestation and its specific cellular localization indicate that this transporter may play an important role in materno-fetal cholesterol transfer.