Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 8.139
Filtrar
4.
Endocrinol. diabetes nutr. (Ed. impr.) ; 66(7): 434-442, ago.-sept. 2019. ilus, graf, tab
Artigo em Inglês | IBECS | ID: ibc-182863

RESUMO

Background: Non-alcoholic fatty liver disease (NAFLD), a condition that leads to fibrosis, is caused by intake of very high-fat diets (HFDs). However, while the negative impact on the liver of these diets has been an issue of interest, systematic research on the effect of HFDs are lacking. Objective: To characterize the overall impact of HFDs on both molecular and morphological signs of liver remodeling. Methods: A study was conducted on male C57BL/6J mice to assess the effect of 4- and 8-week HFDs (60% kcal from fat) on (I) liver steatosis and fibrosis, and (II) expression of factors involved in inflammation and angiogenesis. Results: After an 8-week HFD, vascular endothelial growth factor type-2 receptor (VEGF-R2) and fatty acid translocase/trombospondin-1 receptor (CD36) were overexpressed in liver tissue of mice given HFDs. These changes suggest impaired liver angiogenesis and occurred together with (I) increased GPR78-BiP and EIF2alpha phosphorylation, suggesting endoplasmic reticulum stress, (II) induction of Col1a1 gene expression, a marker of fibrosis, and (III) increased CD31 immunolabeling, consistent with active angiogenesis and fibrosis. Conclusion: Our data show that very HFDs promote a rapid inflammatory response, as well as deregulation of angiogenesis, both consistent with development of liver fibrosis


Antecedentes: El hígado graso no alcohólico (HGNA) es una enfermedad hepática que ocasiona fibrosis y se genera por la ingesta de dietas ricas en grasa. Aunque los efectos nocivos de este tipo de dietas son de gran interés, no son muy abundantes los estudios sistemáticos sobre las consecuencias que su consumo puede tener en el hígado. Objetivo: Evaluar los efectos de una dieta rica en grasa sobre el remodelado hepático, tanto a nivel morfológico como molecular. Métodos: Se utilizaron ratones macho C57BL/6J tratados durante 4/8 semanas con una dieta que contenía un 60% de las kilocalorías procedentes de grasa sobre: 1) la aparición de esteatosis y/o fibrosis hepática y 2) la expresión de factores implicados en procesos de inflamación y angiogénesis. Resultados: Tras 8 semanas de dieta se observó un incremento en el receptor del factor de crecimiento vascular endotelial tipo 2 (R2-FCVE) y en la translocasa de ácidos grasos (CD36). Estos cambios, que sugieren que los procesos angiogénicos del hígado están alterados, fueron simultáneos a: 1) un aumento del GPR78-BiP y de la fosforilación de EIF2alpha, marcadores de estrés del retículo endoplásmico, 2) la inducción en la expresión del gen de colágeno Col1a1, indicador de fibrosis y 3) un incremento de células inmunofluorescentes para CD31 compatible con procesos de angiogénesis y de fibrosis. Conclusiones: Nuestros resultados muestran que las dietas con alto contenido en grasa inducen la rápida activación de respuestas inflamatorias, así como alteraciones en la angiogénesis, siendo ambos procesos compatibles con el desarrollo de fibrosis hepática


Assuntos
Animais , Camundongos , Gorduras na Dieta/efeitos adversos , Dieta Hiperlipídica/efeitos adversos , Inibidores da Angiogênese , Fígado Gorduroso/complicações , Fígado Gorduroso/veterinária , Inflamação/complicações , Fatores de Crescimento Endotelial/administração & dosagem , Antígenos CD36 , Fibrose , Projetos de Pesquisa , Western Blotting
5.
Biomed Pharmacother ; 116: 109007, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31170663

RESUMO

It's critical for tube formation and angiogenesis to repair ischemic myocardium or stroke. This study aimed to investigate role of microRNA-126 (miR-126) in tube formation in human umbilical vein endothelial cells (HUVECs) and associated mechanisms. Primary neural stem cells (NSCs) and HUVECs were cultured and transfected with microRNA-126 mimics and miR-126 inhibitor. Cell counting kit-8 (CCK-8) and cell cycle assay were conducted for evaluating NSCs viability. Transwell assay was conducted to observe invasive ability of HUVECs. Quantitative real-time PCR (qRT-PCR) assay was used to examine epidermal growth factor like domain 7 (EGFL7) and miR-126 mRNA both in vitro and animal models. Tube forming capability was evaluated in HUVECs. Dual luciferase assay was performed to evaluate interaction between miR-126 and EGFL7 gene. Western blot assay was used to determine phosphoinositide-3-kinase/protein kinase-B (PI3K/AKT) signaling molecules and EGFL7. The results indicated that miR-126 significantly decreased cell viability, inhibited invasive ability and modulated cell cycle of NSCs compared to miR-NC group (p < 0.05). miR-126 significantly inhibited tube formation of HUVECs compared to miR-NC group (p < 0.05). miR-126 significantly down-regulated EGFL7 mRNA and protein expression compared to miR-NC (p < 0.05). Atorvastatin significantly increased CD34 and enhanced EGFL7 expression in traumatic brain injury (TBI) rats brain tissues compared to Model group (p < 0.05). miR-126 significantly down-regulated and atorvastatin up-regulated PI3K/AKT signaling pathway (p < 0.05). Atorvastatin significantly increased EGFL7 and down-regulated miR-126 expression in TBI rats brain tissues compared to Model group (p < 0.05). miR-126 interacted with and negatively correlated with EGFL7 gene both in vitro and in TBI models. In conclusion, microRNA-126 inhibited tube formation of HUVECs by interacting with EGFL7 and down-regulating PI3K/AKT signaling pathway.


Assuntos
Proteínas de Ligação ao Cálcio/metabolismo , Regulação para Baixo/genética , Família de Proteínas EGF/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , MicroRNAs/metabolismo , Neovascularização Fisiológica , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Animais , Antígenos CD34/metabolismo , Atorvastatina/administração & dosagem , Atorvastatina/farmacologia , Sequência de Bases , Lesões Encefálicas Traumáticas/genética , Lesões Encefálicas Traumáticas/patologia , Proteínas de Ligação ao Cálcio/genética , Ciclo Celular/efeitos dos fármacos , Ciclo Celular/genética , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Regulação para Baixo/efeitos dos fármacos , Família de Proteínas EGF/genética , Fatores de Crescimento Endotelial/metabolismo , Humanos , MicroRNAs/genética , Neovascularização Fisiológica/efeitos dos fármacos , Neovascularização Fisiológica/genética , Células-Tronco Neurais/efeitos dos fármacos , Células-Tronco Neurais/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genética
8.
Mol Cancer ; 18(1): 81, 2019 04 05.
Artigo em Inglês | MEDLINE | ID: mdl-30953521

RESUMO

BACKGROUND: The aberrant expression of long noncoding RNAs (lncRNAs) has recently emerged as key molecules in human cancers; however, whether lncRNAs are implicated in the progression of clear cell renal cell carcinoma (ccRCC) remains unclear. METHODS: Candidate lncRNAs were selected using microarray analysis and quantitative real-time PCR (qRT-PCR) was performed to detect lncRNAs expression in human ccRCC tissues. Overexpression and knocking down experiments in vivo and in vitro were performed to uncover the biological roles of lncRNA-URRCC on ccRCC cell proliferation and invasion. Microarray, chromatin immunoprecipitation, Luciferase reporter assay and western blot were constructed to investigate the molecular mechanisms underlying the functions of lncRNA-URRCC. RESULTS: The microarray analysis and qRT-PCR identified a new lncRNA, URRCC, whose expression is upregulated in RCC samples and associated with poor prognosis, leading to promote ccRCC cell proliferation and invasion. Mechanistically, URRCC enhances the expression of EGFL7 via mediating histone H3 acetylation of EGFL7 promoter, activation of P-AKT signaling, and suppressing P-AKT downstream gene, FOXO3. In return, FOXO3 could inhibit the transcription of URRCC via binding to the special region on the promoter of URRCC. CONCLUSIONS: Our data suggests that targeting this newly identified feed-back loop between LncRNA-URRCC and EGFL7/P-AKT/FOXO3 signaling may enhance the efficacy of existing therapy and potentially imparts a new avenue to develop more potent therapeutic approaches to suppress RCC progression.


Assuntos
Carcinoma de Células Renais/patologia , Neoplasias Renais/patologia , RNA Longo não Codificante/genética , Transdução de Sinais , Animais , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Fatores de Crescimento Endotelial/metabolismo , Proteína Forkhead Box O3/metabolismo , Perfilação da Expressão Gênica , Humanos , Neoplasias Renais/genética , Neoplasias Renais/metabolismo , Camundongos , Metástase Neoplásica , Estadiamento de Neoplasias , Transplante de Neoplasias , Análise de Sequência com Séries de Oligonucleotídeos , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Regulação para Cima
11.
Transl Res ; 207: 19-29, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30620888

RESUMO

Proper placental development is crucial to establish a successful pregnancy. Defective placentation is the major cause of several pregnancy complications, including preeclampsia (PE). We have previously demonstrated that the secreted factor Epidermal Growth Factor-like Domain 7 (EGFL7) is expressed in trophoblast cells of the human placenta and that it regulates trophoblast migration and invasion, suggesting a role in placental development. In the present study, we demonstrate that circulating levels of EGFL7 are undetectable in nonpregnant women, increase during pregnancy and decline toward term. Close to term, circulating levels of EGFL7 are significantly higher in patients affected by PE when compared to normal pregnancies. Consistent with these results, villus explant cultures obtained from placentas affected by PE display increased release of EGFL7 in the culture medium when compared to those from normal placentas. Our results suggest that increased release of placenta-derived EGFL7 and increased circulating levels of EGFL7 are associated with the clinical manifestation of PE.


Assuntos
Fatores de Crescimento Endotelial/sangue , Pré-Eclâmpsia/sangue , Adulto , Endoglina/sangue , Análise Fatorial , Feminino , Humanos , Modelos Logísticos , Análise Multivariada , Fator de Crescimento Placentário/sangue , Gravidez , Análise de Componente Principal , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/sangue
12.
Neoplasma ; 66(2): 187-196, 2019 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-30569717

RESUMO

Melanoma is the main cause of death in patients with skin cancer. While the pathogenesis of cutaneous melanoma is poorly understood, increasing evidence shows that epidermal growth factor (EGF) may be involved. Herein, we tested the hypothesis that down-regulation of EGFL7 inhibits development and progression of human cutaneous melanoma (CM). Initially, we performed immunohistochemical analysis of EGFL7 in 130 specimens and the findings indicated that EGFL7 was highly expressed in CM. The expressions of EGFL7 and Notch signaling pathway-related genes in CM were then measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot assay. In order to assess biological functions of EGFL7 in CM we up-regulated or down-regulated endogenous EGFL7 using EGFL7-OE or shRNA against EGFL7 in the A375 CM cell line. To better understand the pivotal role of Notch signaling pathway in CM, we blocked this pathway in A375 cells by inhibitor treatment. Finally, tumor xenograft in nude mice was performed to test the in vivo tumorigenesis of the transfected A375 cells. While EGFL7 activated the Notch signaling pathway in CM, gain- and loss-of-function studies established that decreased EGFL7 inhibited cell proliferation and promoted apoptosis in A375 cells. Moreover, down-regulated EGFL7 suppressed in vivo tumorigenesis. Most importantly, we determined that down-regulating EGFL7 inhibited CM development by suppressing the Notch signaling pathway. The combined findings define potential roles of decreased EGFL7 as inhibitors of CM development by suppressing the Notch signaling pathway, and EGFL7 may therefore be a novel therapeutic target in cutaneous melanoma patients.


Assuntos
Fatores de Crescimento Endotelial/genética , Inativação Gênica , Melanoma/genética , Receptores Notch/metabolismo , Transdução de Sinais , Neoplasias Cutâneas/patologia , Animais , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Humanos , Melanoma/patologia , Camundongos , Camundongos Nus , Transplante de Neoplasias , Proteínas , Neoplasias Cutâneas/genética
16.
Eur J Histochem ; 62(4)2018 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-30504933

RESUMO

Cholangiocarcinoma (CCA) is an aggressive biliary tract malignancy with limited treatment options and low survival rates. The intrahepatic subtype comprises two forms: mucin-iCCA and mixed-iCCA. Epidermal growth factor-like domain multiple (EGFL7) is overexpressed in less differentiated liver tumors. The aim of this study was to assess the presence of EGFL7 due to its possible role in the growth of CCA. Hematoxylin and Eosin and periodic acid-Schiff staining were used to evaluate the morphological aspects and glycogen deposition. Immunohistochemistry and immunofluorescence were performed to identify the presence of EGFL7 both in tumor sections ex vivo and in appropriate cell lines in culture. We found that EGFL7 is expressed in malignant cholangiocytes of mixed-iCCA and absent in mucin-iCCA. In conclusion the expression of EGFL7 might be useful in the classification of CCA subtypes.


Assuntos
Neoplasias dos Ductos Biliares/metabolismo , Ductos Biliares Intra-Hepáticos/metabolismo , Colangiocarcinoma/metabolismo , Fatores de Crescimento Endotelial/metabolismo , Idoso , Idoso de 80 Anos ou mais , Neoplasias dos Ductos Biliares/patologia , Ductos Biliares Intra-Hepáticos/patologia , Linhagem Celular Tumoral , Colangiocarcinoma/patologia , Epitélio/metabolismo , Epitélio/patologia , Humanos , Pessoa de Meia-Idade
17.
Pregnancy Hypertens ; 14: 115-120, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30527097

RESUMO

INTRODUCTION: Preeclampsia is a severe complication of pregnancy, and likely arises from abnormal placental development in early pregnancy. Persistent placental hypoxia is thought to trigger the release of anti-angiogenic factors into the maternal circulation leading to widespread endothelial dysfunction. Epidermal growth factor-like domain 7 (EGFL7) is a secreted angiogenic factor that may play a key role in the disrupted angiogenesis seen in response to placental hypoxia that characterizes preeclampsia. METHODS: Primary trophoblasts were isolated and cultured in both normoxic and hypoxic conditions. Under hypoxia HIF1α was silenced and EGFL7 mRNA expression was assessed. EGFL7 mRNA expression was measured in placentas obtained from women with early (<34 weeks) and late onset preeclampsia; and in peripheral whole blood maternal samples from women with preeclampsia and gestation matched controls. EGFL7 plasma levels were assessed in plasma from women with preeclampsia, compared to gestation-matched controls. RESULTS: EGFL7 expression was significantly upregulated in primary human trophoblasts cultured in hypoxia (>2-fold, p < 0.0001), however this was not regulated via a HIF1α dependent manner. EGFL7 mRNA expression was not altered in placenta from women with early or late onset preeclampsia. Circulating EGFL7 protein levels were not different in women with severe preeclampsia. In contrast, EGFL7 mRNA expression was increased in maternal blood in women with early onset preeclampsia (∼1.6-fold, p < 0.05). DISCUSSION: EGFL7 mRNA expression is increased with hypoxia in human trophoblast and is increased in the maternal circulation in women with preeclampsia. Further studies aimed at understanding the role and regulation of EGLF7 in the pathophysiology of preeclampsia are required.


Assuntos
Fatores de Crescimento Endotelial/metabolismo , Pré-Eclâmpsia/metabolismo , Trofoblastos/metabolismo , Adulto , Estudos de Casos e Controles , Fatores de Crescimento Endotelial/sangue , Feminino , Expressão Gênica , Humanos , Hipóxia/metabolismo , Placenta/metabolismo , Pré-Eclâmpsia/sangue , Gravidez , Adulto Jovem
18.
Biomed Res ; 39(6): 287-294, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30531158

RESUMO

Peripheral nerve injury has been suggested to up-regulate mRNA for the vascular endothelial growth factor (VEGF) which enhances nerve regeneration. VEGF is known to regulate angiogenesis by binding with a specific receptor, the vascular endothelial growth factor receptor (VEGFR). However, little is known about the involvement of VEGF-VEGFR signaling in the nerve regeneration at early stages though previous studies contained a lengthy observation. The present study examined that relationship between angiogenesis and peripheral nerve regeneration at the early stage after nerve transection by focusing on the chronological changes in the expression patterns of VEGF-VEGFR signaling. This study used our previously reported experimental model for nerve regeneration following the transection of the inferior alveolar nerve (IAN) in mice. In a double staining of PGP9.5 and CD31, respective markers for the nerve fibers and endothelial cells, CD31 immunoreactions first appeared in the injury site on postoperative (PO) day 2 when the transected nerve fibers had not been re-connected. The most intense immunoreaction for CD31 was found around the regenerating nerve fibers extending from the proximal stump on PO day 3, but it gradually lessened to disappear by PO day 7. The expression patterns of VEGFR1 and VEGFR2 showed similar chronological changes through the observation periods, with most intense immunoreaction found on PO day 3. Western blotting of total protein extracted from the injury site demonstrated the clear bands for VEGF-A and VEGF-B on PO day 2, indicating a time lag for the expression of ligands and receptors. A local administration of antibody to VEGF-A inhibited the elongation of the nerve fibers from the proximal stump. Furthermore, this administration of VEGF-A antibody inhibited the expression of CD31 in the gap between proximal and distal stumps. These results indicated that a nerve injury initiates productions in VEGF-A and VEFG-B, followed with the expression of VEGFR1 and VEGFR2 at early stages after the nerve injury. Taken these findings together, it is reasonable to postulate that immediate response of VEGF-VEGFR signaling to nerve injury plays a crucial role in local angiogenesis, resulting in a trigger for the regeneration of the nerve fibers in mouse IAN.


Assuntos
Fatores de Crescimento Endotelial/metabolismo , Regeneração Nervosa , Transdução de Sinais , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Células Endoteliais/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fibras Nervosas , Molécula-1 de Adesão Celular Endotelial a Plaquetas/antagonistas & inibidores , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Ubiquitina Tiolesterase/metabolismo , Fator B de Crescimento do Endotélio Vascular/metabolismo , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
19.
INSPILIP ; 2(2): 1-18, jul.-dic. 2018.
Artigo em Espanhol | LILACS | ID: biblio-981647

RESUMO

El objetivo de la investigación fue establecer el modelo de expresión de factor de crecimiento endotelial (VEGF) durante la foliculogénesis en el ovario humano. La investigación fue aprobada por el Comité de Éticade la Facultad de Medicina de la Universidad del Zulia. Se recogieron muestras de tejido ovárico de mujeres en edad reproductiva sometidas a cirugía ginecológica por enfermedad benigna. Las células de la granulosa luteinizada y el líquido folicular se obtuvieron de mujeres sometidas a la recuperación de oocitos para la fertilización in vitro. Utilizando inmunohistoquímica, se localizó el VEGF en las células de la granulosa y la teca de los folículos antrales y en las células luteínicas del cuerpo lúteo. También se identificó en el líquido folicularhumano. El VEGF y el ARN mensajero se identificaron por inmunohistoquímica en la granulosa luteinizada del ovario. La hibridación in situ demostró la expresión del ARN mensajero del VEGF en las células de luteína del cuerpo lúteo. Se concluye que las células de la granulosa y la teca de los folículos antrales y las células de luteína del cuerpo lúteo son fuentes de VEGF en el ovario humano. Su expresión es paralela al patrón de la angiogénesis ovárica y su presencia en el líquido folicular humano sugiere un posible papel en la formación de este.


The objective of the research was to establish the expression model of endothelial growth factor during folliculogenesis in the human ovary. The research was approved by the Ethics Committee of the Faculty of Medicine of LaUniversity of Zulia. Ovarian tissue samples were collected from women of reproductive age who underwent gynecological surgery for benign disease. Luteinized granulosa cells and follicular fluid were obtained from women undergoing oocyte recovery for in vitro fertilization. Using immunohistochemistry, VEGF was localized in the granulosa and teak cells of the antral follicles and in the luteal cells of the corpus luteum. It was also identified in the human follicular fluid. VEGF and messenger RNA were identified by immunohistochemistry in the luteinized granulosa of the ovary. In situ hybridization demonstrated the expression of VEGF messenger RNA in the lutein cells of the corpus luteum. It is concluded that the granulosa and teak cells of the antral follicles and the lutein cells of the corpus luteum are sources of VEGF in the human ovary. Its expression is parallel to the pattern of ovarian angiogenesis and its presence in the human follicular fluid suggests a possible role in the formation of it.


Assuntos
Humanos , Feminino , Fertilização In Vitro , Fatores de Crescimento Endotelial , Folículo Ovariano , Hibridização Genética , Venezuela
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA