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BACKGROUND: and purpose: Ghrelin is the endogenous ligand of the growth hormone secretagogue receptor (GHS-R1). Ghrelin, and GHS-R1, may have a role in placental growth and function, and its unacylated form desacylghrelin (DAG) could be involved in fetal growth. Nevertheless, the effects of DAG on placental function, and the receptor involved in its actions, remain to be determined. We aimed to investigate the effect of DAG in placental BeWo cells viability, proliferation, differentiation, and GSH-R1 expression. METHODS: BeWo cells, a human trophoblast cell line, was cultured with 3 nM DAG during 12, 24, 48, and 72 h. Cell viability, proliferation, differentiation (assessed by human Chorionic Gonadotropin quantification), and GSH-R1 expression were analyzed. To evaluate the mechanism of DAG effect on GSH-R1, 30 nM receptor antagonist ([D-Lys3]-GHRP-6) was added alone or in combination with 3 nM DAG during 12 h and 24 h. RESULTS: DAG has no effect on cell proliferation or viability, but it has an inhibitory effect on cell differentiation. DAG had a stimulatory effect on GSH-R1 expression at 12 and 24 h (p = 0.029 and p = 0.025, respectively). On the contrary, culture with 48 h DAG inhibits GSH-R1 expression compared to the control (p = 0.005), while GSH-R1 antagonist inhibited the effect of DAG on GSH-R1 expression. DAG also reduces intracellular (p = 0.020) and secreted (p = 0.011) hCG concentration in BeWo cells. CONCLUSION: DAG increases GHS-R1 expression, potentially mediated through GHS-R1 itself. DAG may also inhibit placental BeWo cell differentiation, suggesting a possible role of DAG in placental and fetal physiology.

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The respiratory syncytial virus (RSV) is the main pathogen associated with upper respiratory tract infections during early childhood. Vertical transmission of this virus has been suggested in humans, based on observations recorded during animal studies that revealed an association of RSV with persistent structural and functional changes in the developing lungs of the offspring. However, human placentas have not yet been evaluated for susceptibility to RSV infection. In this study, we examined the capacity of RSV to infect a human trophoblast model, the BeWo cell line. Our results suggest that BeWo cells are susceptible to RSV infection since they allow RNA viral replication, viral protein translation, leading to the production of infectious RSV particles. In this report, we demonstrate that a human placenta model system, consisting of BeWo cells, is permissive to RSV infection. Thus, the BeWo cell line may represent a useful model for studies that aim to characterize the events of a possible RSV infection at the human maternal-fetal interface.

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Las células estromales son las células más abundantes presentes en la decidua y juegan un papelmuy importante durante la implantación, la nutrición fetal y el mantenimiento del embarazo. Losprocedimientos que se han descrito para el aislamiento de células estromales requieren el uso de muchosanticuerpos monoclonales ya que hay contaminación con otros tipos celulares en la decidua y ademásalgunos marcadores características de células estromales, muestran variabilidad en los diferentes díasde la gestación. En este estudio se estandarizó un procedimiento de aislamiento por digestión enzimática,gradiente de densidad y adherencia al plástico y se caracterizaron las células estromales murinas porexclusión de marcadores que se expresan en macrófagos (F4-80), células epiteliales y trofoblasto(citoqueratina-7), obteniéndose un 98% de células negativas para estos marcadores que correspondería alas células estromales. Esta técnica de aislamiento permite obtener células estromales con métodos menoscostosos y altamente eficientes que facilita el acceso a un modelo celular de gran utilidad en el estudio dela fisiología de la gestación en diferentes especies.

Stromal cells are the most abundant cell population present in decidual tissue; they are involved inkey processes during embryo implantation, fetal nutrition and the pregnancy maintenance. Described procedures for stromal cells isolation require the use of many monoclonal antibodies due to contaminationwith another cell types in the decidua; besides, some markers of stromal cells show variability during thedays of gestation. In this study, we standardized a procedure for isolation by enzymatic digestion, densitygradient and adherence to plastic. Murine stromal cells were characterized by exclusion of markers thatare expressed in macrophages (F4-80), epithelial cells and trophoblast (cytokeratin-7), yielding a 98% ofnegative cells for these markers that correspond to stromal cells. This isolation procedure permits to obtainstromal cells with less expensive and high efficiency methods that provide a useful cellular model to studythe physiology of gestation in different species.

As células estromales são as células mais abundantes presentes na decídua e tem um papel muitoimportante durante a implementação, a nutrição fetal e a manutenção da gravidez. Os procedimentosdescritos para o isolamento das células estromales necessitam o uso de muitos anticorpos monoclonais jáque há contaminação com outros tipos celulares na decídua e demais alguns marcadores característicosdas células estromales que tem variabilidade nos diferentes dias de gestação. No presente trabalho foiestandardizado um procedimento de isolamento por digestão enzimática, gradiente de densidade eaderência ao plástico e foram caracterizados as células estromales murinas por exclusão de marcadoresque expressão em macrófagos (F4-80), células epiteliais e trofoblasto (citoqueratina-7), obtendo-se um98% de células negativas para estes marcadores que corresponderia às células estromales. Esta técnicade isolamento permite obter células estromales com métodos menos custosos e altamente eficientes quefacilita o aceso a um modelo celular de grande utilidade no estudo da fisiologia da gestação em diferentesespécies.

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