RESUMO
Disruption of the blood-brain barrier (BBB) is central in the pathophysiology of acute cerebral complications in women who have preeclampsia. Underling mechanisms are unclear. Using female human brain endothelial cells as an in vitro model of BBB, we show that plasma of women with preeclampsia increases cell apoptosis and permeability via activation of the vascular endothelial growth factor receptor 2 (VEGFR2). Since plasma of women with preeclampsia also enhanced VEGFR2 phosphorylation in the tyrosine 951 but decreased phosphorylation at the tyrosine 1175, we propose the former would be the more likely active form of VEGFR2 responsible for BBB alterations.
Assuntos
Pré-Eclâmpsia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Apoptose , Barreira Hematoencefálica/metabolismo , Permeabilidade Capilar , Células Endoteliais/metabolismo , Feminino , Humanos , Fosforilação , Pré-Eclâmpsia/metabolismo , Gravidez , Tirosina/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Resumen La crisis sanitaria por la pandemia de COVID-19, enfermedad generada por la infección con el SARS-CoV-2, ha llevado a la pérdida de más de 25,000 vidas en Chile, con más de 370,000 mujeres entre 15 y 44 años con infección confirmada. Se ha reportado una mayor vulnerabilidad de las mujeres gestantes en cuanto a desarrollar cuadros de COVID-19 graves o críticos, con un aumento de la incidencia de resultados obstétricos y perinatales adversos. Es relevante considerar que un alto porcentaje de las gestantes infectadas con SARS-CoV-2 son asintomáticas, lo cual nos pone en alerta en cuanto a que ciertos efectos del virus durante la gestación podrían no ser evidentes para la observación clínica. Se ha demostrado la presencia de SARS-CoV-2 en la placenta, asociándose la infección placentaria con alteraciones vasculares que podrían afectar el flujo útero-placentario. Por otro lado, la transmisión vertical al feto parece que es poco frecuente, pero factible. Se resumen las evidencias disponibles hasta el momento sobre los principales efectos de la COVID-19 en la gestación, con énfasis en los estudios sobre los efectos de la infección por SARS-CoV-2 en la placenta. El objetivo es relevar el tema, destacando que son diversas las preguntas que necesitan ser abordadas, considerando el impacto que esta pandemia podría tener sobre la salud gestacional.
Abstract In Chile, the COVID-19 pandemic, a disease induced by infection with SARS-CoV-2, has caused more than 25,000 deaths. More than 370,000 women between 15 and 44 years have been detected with the infection. The greater vulnerability of pregnant women has been reported, mainly related to a higher risk for severe or critical COVID-19, with an increased incidence of adverse obstetrics and perinatal outcomes. It is relevant to consider that a high percentage of pregnant women infected with SARS-CoV-2 are asymptomatic for COVID-19, which indicates that specific effects of the virus during pregnancy may not be evident from clinical observation. The presence of SARS-CoV-2 in the placenta has been demonstrated, associating placental infection with vascular alterations that could affect utero-placental flow. On the other hand, vertical transmission to the fetus is rare but feasible. This manuscript summarizes the evidence available to date on the main effects of COVID-19 in pregnancy, emphasizing studies about the impact of SARS-CoV-2 in the placenta. This review aims to promote this issue, highlighting that several questions need to be addressed, considering the effect this pandemic could have on gestational health.
Assuntos
Humanos , Feminino , Gravidez , Placenta/virologia , Complicações Infecciosas na Gravidez/epidemiologia , COVID-19/complicações , Transmissão Vertical de Doenças Infecciosas , Pandemias , SARS-CoV-2/patogenicidade , COVID-19/fisiopatologia , COVID-19/transmissãoRESUMO
Estrogenic steroids and adenosine A2A receptors promote the wound healing and angiogenesis processes. However, so far, it is unclear whether estrogen may regulate the expression and pro-angiogenic activity of A2A receptors. Using in vivo analyses, we showed that female wild type (WT) mice have a more rapid wound healing process than female or male A2A-deficient mice (A2AKO) mice. We also found that pulmonary endothelial cells (mPEC) isolated from female WT mice showed higher expression of A2A receptor than mPEC from male WT mice. mPEC from female WT mice were more sensitive to A2A-mediated pro-angiogenic response, suggesting an ER and A2A crosstalk, which was confirmed using cells isolated from A2AKO. In those female cells, 17ß-estradiol potentiated A2A-mediated cell proliferation, an effect that was inhibited by selective antagonists of estrogen receptors (ER), ERα, and ERß. Therefore, estrogen regulates the expression and/or pro-angiogenic activity of A2A adenosine receptors, likely involving activation of ERα and ERß receptors. Sexual dimorphism in wound healing observed in the A2AKO mice process reinforces the functional crosstalk between ER and A2A receptors.
Assuntos
Estradiol/farmacologia , Receptor alfa de Estrogênio/genética , Receptor beta de Estrogênio/genética , Neovascularização Fisiológica/efeitos dos fármacos , Receptor A2A de Adenosina/genética , Ferimentos Penetrantes/genética , Adenosina/análogos & derivados , Adenosina/farmacologia , Animais , Proliferação de Células/efeitos dos fármacos , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Receptor alfa de Estrogênio/antagonistas & inibidores , Receptor alfa de Estrogênio/metabolismo , Receptor beta de Estrogênio/antagonistas & inibidores , Receptor beta de Estrogênio/metabolismo , Feminino , Regulação da Expressão Gênica , Pulmão/citologia , Pulmão/metabolismo , Masculino , Camundongos , Camundongos Knockout , Neovascularização Fisiológica/genética , Fenetilaminas/farmacologia , Pirazóis/farmacologia , Pirimidinas/farmacologia , Receptor Cross-Talk , Receptor A2A de Adenosina/metabolismo , Fatores Sexuais , Transdução de Sinais , Cicatrização/efeitos dos fármacos , Cicatrização/genética , Ferimentos Penetrantes/tratamento farmacológico , Ferimentos Penetrantes/metabolismo , Ferimentos Penetrantes/patologiaRESUMO
We aim to investigate the role of A2A receptor in peritonitis-related sepsis by injection of a fecal solution (FS) as a model of polymicrobial infection. C57/black J6 wild-type (WT) and A2A-deficient mice (A2AKO) were exposed to sepsis induced by intraperitoneal injection of a FS (FS-induced peritonitis) or instead was injected with saline buffer (Sham). Survival rate and sepsis score were measured up to 48 h. The presence of bacteria in tissue homogenates was analyzed. Telemetry and speckle laser Doppler were used for systemic blood pressure and peripheral blood perfusion analysis, respectively. Histological analysis and identification of active caspase 3 were performed in selected organs, including the liver. The survival rate of A2AKO mice exposed to FS-induced peritonitis was significantly higher, and the sepsis score was lower than their respective WT counterpart. Injection of FS increases (50 to 150 folds) the number of colonies forming units in the liver, kidney, blood, and lung in WT mice, while these effects were significantly attenuated in A2AKO mice exposed to FS-induced peritonitis. A significant reduction in both systolic and diastolic blood pressure, as well as in the peripheral perfusion was observed in WT and A2AKO mice exposed to FS-induced peritonitis. Although, these last effects were significantly attenuated in A2AKO mice. Histological analysis showed a large perivascular infiltration of polymorphonuclear in the liver of WT and A2AKO mice exposed to FS-induced peritonitis, but again, this effect was attenuated in A2AKO mice. Finally, high expression of active caspase 3 was found only in the liver of WT mice exposed to FS-induced peritonitis. The absence of the A2A receptor increases the survival rate in mice exposed to polymicrobial sepsis. This outcome was associated with both hemodynamic compensation and enhanced anti-bacterial response.
Assuntos
Peritonite/metabolismo , Receptor A2A de Adenosina/metabolismo , Sepse/metabolismo , Animais , Pressão Sanguínea/fisiologia , Modelos Animais de Doenças , Camundongos , Camundongos Knockout , Peritonite/genética , Peritonite/microbiologia , Peritonite/mortalidade , Receptor A2A de Adenosina/genética , Sepse/genética , Sepse/mortalidade , Taxa de SobrevidaRESUMO
We aimed to compare the cardiorespiratory, metabolic, and perceptual responses to high- and moderate-intensity eccentric cycling versus moderate-intensity concentric cycling in chronic obstructive pulmonary disease (COPD) patients. Ten patients with moderate COPD (forced expiratory volume in 1 s (FEV1) = 68.6% ± 20.4% of predicted; 68.3 ± 9.1 years) performed 30 min of moderate-intensity concentric (CONC-M: 50% maximum workload; Wmax), moderate-intensity eccentric (ECC-M: 50% Wmax), and high-intensity eccentric (ECC-H: 100% Wmax) cycling. Average power output, oxygen consumption (VÌO2), minute ventilation (VE), respiratory frequency (fR), oxygen saturation (SpO2), heart rate (HR), systolic and diastolic blood pressure (SBP and DBP), rate of perceived exertion (RPE), and dyspnea were measured during cycling. Compared with CONC-M, lower VÌO2 (-52% ± 14%), VE (-47% ± 16%), fR (-21% ± 14%), HR (-14% ± 16%), SBP (-73% ± 54%), RPE (-36% ± 26%), and dyspnea (-41% ± 37%) were found during ECC-M. During ECC-H, a similar metabolic demand to CONC-M was found. However, average power output was 117% ± 79% higher during ECC-H. Eccentric cycling can be safely performed by COPD patients and induced lower cardiorespiratory, metabolic, and perceptual responses than concentric exercise when performed at the same workload. Novelty Moderate- and high-intensity eccentric cycling can be performed by COPD patients. Moderate-intensity eccentric cycling showed lower cardiorespiratory, metabolic, and perceptual demand than concentric cycling at the same workload in COPD patients. Even at double workload, eccentric cycling induces lower cardiorespiratory, metabolic, and perceptual demand than moderate-intensity concentric cycling.
Assuntos
Exercício Físico , Esforço Físico , Doença Pulmonar Obstrutiva Crônica/metabolismo , Idoso , Pressão Sanguínea , Frequência Cardíaca , Humanos , Pessoa de Meia-Idade , Consumo de OxigênioRESUMO
Placentas from gestational diabetes mellitus (GDM) are often hypervascularized; however, participation of vascular endothelial growth factor (VEGF) and its receptors in this placental adaptation is unclear. We aimed to test whether changes in phosphorylation of tyrosine 951 or tyrosine 1175 (pY951 or pY1175) of the vascular endothelial growth factor receptor 2 (KDR) are associated with the proangiogenic state observed in placentas from GDM. We obtained placental samples from women with normal pregnancies (n = 24) or GDM (n = 18). We measured the relative expression of markers for endothelial cell number (CD31, CD34), VEGF, vascular endothelial growth factor receptor 1 (Flt-1), KDR, pY951 and pY1175 of KDR in placental homogenate. Immunohistochemistry of placental blood vessels were performed using CD34. Proliferation and migration of human umbilical vein endothelial cells (HUVEC) obtained from normal pregnancy and GDM were determined in absence or presence of conditioned medium (CM) harvested from GDM or normoglycemic HUVEC cultures. GDM was associated with more CD31 and CD34 protein compared to normal pregnancy. High number, but reduced area of placental blood vessels was found in GDM. Reduced Flt-1 levels (mRNA and protein) are associated with reduced KDR mRNA, but higher KDR protein levels in placentas from GDM. No significant changes in Y951-or Y1175-phosphorylation of KDR in placentas from GDM were found. GDM did not alter proliferation of HUVECs, but enhanced migration. Conditioned medium harvested from GDM HUVEC cultures enhanced KDR protein amount, tube formation capacity and cell migration in HUVEC isolated from normoglycemic pregnancies. The data indicate that GDM is associated with reduced expression of Flt-1 but high pro-migratory activation of KDR reflecting a proangiogenic state in GDM.
Assuntos
Movimento Celular , Diabetes Gestacional/metabolismo , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Adulto , Antígenos CD34/genética , Antígenos CD34/metabolismo , Biomarcadores/metabolismo , Células Cultivadas , Diabetes Gestacional/diagnóstico , Feminino , Células Endoteliais da Veia Umbilical Humana/metabolismo , Células Endoteliais da Veia Umbilical Humana/fisiologia , Humanos , Recém-Nascido , Masculino , Placenta/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Gravidez , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/genéticaRESUMO
The underlying molecular mechanisms involve in the regulation of the angiogenic process by insulin are not well understood. In this review article, we aim to describe the role of insulin and insulin receptor activation on the control of angiogenesis and how these mechanisms can be deregulated in human diseases. Functional expression of insulin receptors and their signaling pathways has been described on endothelial cells and pericytes, both of the main cells involved in vessel formation and maturation. Consequently, insulin has been shown to regulate endothelial cell migration, proliferation, and in vitro tubular structure formation through binding to its receptors and activation of intracellular phosphorylation cascades. Furthermore, insulin-mediated pro-angiogenic state is potentiated by generation of vascular growth factors, such as the vascular endothelial growth factor, produced by endothelial cells. Additionally, diseases such as insulin resistance, obesity, diabetes, and cancer may be associated with the deregulation of insulin-mediated angiogenesis. Despite this knowledge, the underlying molecular mechanisms need to be elucidated in order to provide new insights into the role of insulin on angiogenesis.
RESUMO
Para la enseñanza de la Anatomía siempre se está en búsqueda de alternativas para la conservación y presentación de preparados anatómicos, realizando variaciones de diferentes técnicas, que permitan presentar especímenes con fines didácticos, para lograr un mejor aprendizaje, comprensión y motivación por la Morfología. El objetivo de este trabajo fue utilizar una combinación de osteotecnia más la conservación de músculos, en un montaje único que permite estudiar en una mitad el esqueleto, y en la otra, la musculatura superficial de un Canis lupus familiaris. Se utilizó un cadáver del especimen fijado por congelación, luego se realizó una limpieza general, extracción de vísceras y retiro de tegumentos previo al corte sagital. En la mitad derecha se realizó la limpieza de los huesos mediante ebullición combinada con métodos mecánicos y posteriormente, el desengrasado y blanqueado de ellos previo al ensamble del esqueleto. Para la conservación de músculos la mitad izquierda del cuerpo fue sumergida en "solución fijadora conservadora chilena" libre de formol, por 31 días, tras lo cual se realizó la disección de fascias y tejido graso para delimitar músculos superficiales. Una vez tratado cada segmento, se procedió al montaje combinado de las mitades en posición anatómica del espécimen, usando suturas desde craneal a caudal, sobre pedestales. La técnica utilizada de montaje combinado proporciona un material anatómico-didáctico flexible, de bajo costo y escasa toxicidad, con la potencialidad de permitir reforzar de manera interactiva el aprendizaje significativo de los estudiantes, facilitando la observación y estudio de la topografía de huesos y músculos.
The teaching of anatomy is always in search of alternatives for the preservation and presentation of anatomical preparations, doing variations of different techniques that allow to present specimens with didactic goals so as to get a better learning, comprehension and motivation for morphology. The purpose of this study was to use a combination of osteotecnia and the preservations of muscles in a unique assembly allowing to study half of the skeleton and, in the other, the superficial muscles of a Canis lupus familiaris. A corpse of the specimen was used, fixed by freezing; then, a general cleaning was carried out, extraction of viscera and removal of teguments before the sagittal cut. On the right half, using boiling combined with mechanical methods and, subsequently, their degreasing and bleaching, prior to the skeleton's assembly carried out the cleaning of the bones. For the preservation of muscles, the left half of the body was submerged in a "Chilean preserving and fixative solution" free from formaldehyde for 31 days, following the dissection of fascia and fatty tissue to delimit superficial muscles. Once each segment is treated, the combined assembly of the halves is made in anatomical position of the specimen, using stitches from cranial to caudal, over pedestals. The utilized technique of combined assembly provides a flexible didactic-anatomical material, of low-cost and low-toxicity, with the potentiality to allow reinforcing in an interactive way the meaningful learning of the students, making the observation easier as well as the study of topography of bones and muscles.
Assuntos
Animais , Anatomia/métodos , Cães/anatomia & histologia , Músculos/anatomia & histologia , Preservação de Tecido/métodos , Modelos Anatômicos , Materiais de EnsinoRESUMO
We aim to investigate whether A2A/nitric oxide-mediated regulation of vascular endothelial growth factor (VEGF) expression is impaired in feto-placental endothelial cells from late-onset pre-eclampsia. Cultures of human umbilical vein endothelial cells (HUVECs) and human placental microvascular endothelial cells (hPMECs) from normal and pre-eclamptic pregnancies were used. Assays by using small interference RNA (siRNA) for A2A were performed, and transfected cells were used for estimation of messenger RNA (mRNA) levels of VEGF, as well as for cell proliferation and angiogenesis in vitro. CGS-21680 (A2A agonist, 24 h) increases HUVEC and hPMEC proliferation in a dose response manner. Furthermore, similar to CGS-21680, the nitric oxide donor, S-nitroso-N-acetyl-penicillamine oxide (SNAP), increased cell proliferation in a dose response manner (logEC50 10-9.2 M). In hPMEC, CGS-21680 increased VEGF protein levels in both normal (â¼1.5-fold) and pre-eclamptic pregnancies (â¼1.2-fold), an effect blocked by the A2A antagonist, ZM-241385 (10-5 M) and the inhibitor of NO synthase, N ω-nitro-L-arginine methyl ester hydrochloride (L-NAME). Subsequently, SNAP partially recovered cell proliferation and in vitro angiogenesis capacity of cells from normal pregnancies exposed to siRNA for A2A. CGS-21680 also increased (â¼1.5-fold) the level of VEGF mRNA in HUVEC from normal pregnancies, but not in pre-eclampsia. Additionally, transfection with siRNA for A2A decrease (â¼30 %) the level of mRNA for VEGF in normal pregnancy compared to untransfected cells, an effect partially reversed by co-incubation with SNAP. The A2A-NO-VEGF pathway is present in endothelium from microcirculation and macrocirculation in both normal and pre-eclamptic pregnancies. However, NO signaling pathway seems to be impaired in HUVEC from pre-eclampsia.
Assuntos
Endotélio Vascular/metabolismo , Pré-Eclâmpsia/metabolismo , Receptor A2A de Adenosina/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adenosina/análogos & derivados , Adenosina/farmacologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Endotélio Vascular/efeitos dos fármacos , Feminino , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Óxido Nítrico/metabolismo , Fenetilaminas/farmacologia , Gravidez , Transdução de Sinais/efeitos dos fármacosRESUMO
Preeclampsia is a syndrome characterized by hypertension during pregnancy, which is a leading cause of morbidity and mortality in both mother and newborn in developing countries. Some advances have increased the understanding of pathophysiology of this disease. For example, reduced utero-placental blood flow associated with impaired trophoblast invasion may lead to a hypoxic placenta that releases harmful materials into the maternal and feto-placental circulation and impairs endothelial function. Identification of these harmful materials is one of the hot topics in the literature, since these provide potential biomarkers. Certainty, such knowledge will help us to understand the miscommunication between mother and fetus. In this review we highlight how placental extracellular vesicles and their cargo, such as small RNAs (i.e., microRNAs), might be involved in endothelial dysfunction, and then in the angiogenesis process, during preeclampsia. Currently only a few reports have addressed the potential role of endothelial regulatory miRNA in the impaired angiogenesis in preeclampsia. One of the main limitations in this area is the variability of the analyses performed in the current literature. This includes variability in the size of the particles analyzed, and broad variation in the exosomes considered. The quantity of microRNA targets genes suggest that practically all endothelial cell metabolic functions might be impaired. More studies are required to investigate mechanisms underlying miRNA released from placenta upon endothelial function involved in the angiogenenic process.
RESUMO
To investigate the functionality of A2B adenosine receptor (A2BAR) and the nitric oxide (NO) and vascular endothelial growth factor (VEGF) signaling pathway in the endothelial cell proliferation/migration during preeclampsia, we used human umbilical vein endothelial cells (HUVECs) isolated from normal pregnancies (n = 15) or pregnancies with preeclampsia (n = 15). Experiments were performed in presence or absence of the nonselective adenosine receptor agonist NECA, the A2BAR selective antagonist MRS-1754, and the nitric oxide synthase (NOS) inhibitor L-NAME. Results indicated that cells from preeclampsia exhibited a significant higher protein level of A2BAR and logEC50 for NECA-mediated proliferation than normotensive pregnancies. The stimulatory effect of NECA (10 µM, 24 h) on cell proliferation was prevented by MRS-1754 (5 nM) coincubation only in cells from normotensive pregnancies. Nevertheless, L-NAME (100 µM, 24 h) reduced the NECA-induced cell proliferation/migration in HUVEC from normal pregnancy; however in preeclampsia only NECA-induced cell proliferation was reduced by L-NAME. Moreover, NECA increased protein nitration and abundance of VEGF in cells from normal pregnancy and effect prevented by MRS-1754 coincubation. Nevertheless, in preeclampsia NECA did not affect the protein level of VEGF. In conclusion HUVECs from preeclampsia exhibit elevated protein level of A2BAR and impairment of A2BAR-mediated NO/VEGF signaling pathway.
Assuntos
Movimento Celular , Proliferação de Células , Endotélio Vascular/metabolismo , Feto/metabolismo , Pré-Eclâmpsia/metabolismo , Receptor A2B de Adenosina/metabolismo , Acetamidas/farmacologia , Adenosina-5'-(N-etilcarboxamida)/farmacologia , Adulto , Antineoplásicos/farmacologia , Endotélio Vascular/patologia , Feminino , Feto/patologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Óxido Nítrico/metabolismo , Pré-Eclâmpsia/patologia , Gravidez , Purinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Fator A de Crescimento do Endotélio VascularRESUMO
El Neurocitoma Central (NC) es un tumor del SNC infrecuente, de estirpe neuronal, frecuentemente intraventricular, que generalmente afecta adultos jòvenes, tiene crecimiento lento y que al momento del diagnóstico tiene con frecuencia un volumen considerable. Su comportamiento es poco agresivo y una exéresis quirúrgica conservadora permite mejorar sustancialmente la calidad y espectativa vital. Presentamos aquí dos casos clínicos de pacientes con cuadros clínicos compatibles a los decritos en la literatura. Se realizaron estudios inmunohistoquímicos de las lesiones que confirman el diagnóstico.
Central Neurocytoma (CN) it's a rare Central Nervous System Tumor, derivated of the neuron, frequently intraventricular, it generally affects young adults, has a slow pattern of growth and at diagnosis is frequently voluminous. It's a less aggressive kind of tumor and a conservative surgery exeresis allows a better quality and expectative of life. We present two cases of patients with similar clinical presentation with the descriptions find in the literature and with histologyc and immunohistochemistry studies that confirms the diagnosis.