RESUMEN
Background: The brain is an organ that serves as the center of the nervous system in all vertebrate and most invertebrate animals. Aim: The study examined the expression of Neuroglobin (Ngb) and Hypoxia-inducible factor-1α (Hif-1α) in adult and young yak brain tissues, and provided researchers with meaningful insight into the anatomy, physiology, and biochemistry of this mammal. Method: The study employed immunohistochemistry (IHC), quantitative real-time PCR (qRT-PCR), and Western blot (WB) to obtain the results. Results: Ngb and Hif-1α were significantly (P<0.05) expressed in the cerebellar cortex, piriform lobe, medulla, and corpus callosum of the adult yak while in the young yak brain tissues, the protein expressions were significantly found in the white matter of the cerebellum, pineal gland, corpus callosum, and cerebellar cortex. The Ngb and Hif-1α expression showed similarities and differences. This may have resulted from similar animal species, source of nutrition, age factors, brain size, emotional activities, and communication. The findings documented that Ngb and Hif-1α are commonly expressed in various adult and young yak brain tissues. Multiple roles in the brain tissues of the adult and young yaks are involved in the expression and distribution and are proposed to play a significant role in the adaptation of the yak to the high altitude environment. Conclusion: This study provides meaningful data to understand the adaptive mechanism to hypoxia and recommended researchers to expand on the adaptive mechanism and brain tissues that are not recorded.
Contexto: O cérebro é um órgão que funciona como o centro do sistema nervoso em todos os animais vertebrados e na maioria dos invertebrados. Objetivo: O estudo examinou a expressão de neuroglobina (Ngb) e fator-1α indutível por hipóxia (Hif-1α) em tecidos cerebrais de iaques adultos e jovens e forneceu aos pesquisadores uma visão significativa da anatomia, fisiologia e bioquímica desse mamífero. Método: O estudo utilizou imuno-histoquímica (IHC), PCR quantitativo em tempo real (qRT-PCR) e western blot (WB) para a obtenção dos resultados. Resultados: Ngb e Hif-1α foram significativamente (P < 0,05) expressos no córtex cerebelar, lobo piriforme, medula e corpo caloso do iaque adulto, enquanto nos tecidos cerebrais do iaque jovem as expressões proteicas foram encontradas significativamente na substância branca do cerebelo, glândula pineal, corpo caloso e córtex cerebelar. A expressão de Ngb e Hif-1α apresentou semelhanças e diferenças. Isso pode ter resultado de espécies animais semelhantes, fonte de nutrição, fatores de idade, tamanho do cérebro, atividades emocionais e comunicação. Os resultados documentaram que o Ngb e o Hif-1α são comumente expressos em vários tecidos cerebrais de iaques adultos e jovens. Múltiplos papéis nos tecidos cerebrais de iaques adultos e jovens estão envolvidos na expressão e distribuição e são propostos para desempenhar um papel significativo na adaptação do iaque ao ambiente de alta altitude. Conclusão: Este estudo fornece dados significativos para compreender o mecanismo adaptativo à hipóxia e recomendou que os pesquisadores expandissem o mecanismo adaptativo e os tecidos cerebrais que não foram registrados.
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Animales , Adulto Joven , Adulto , Bovinos , Bovinos , Cerebro/anatomía & histología , Cerebro/fisiología , Factor 1 Inducible por Hipoxia/análisis , Fenómenos Bioquímicos , Neuroglobina/análisisRESUMEN
The telencephalon refers to the most highly developed and anterior part of the forebrain, consisting mainly of the cerebral hemispheres. The study determined Neuroglobin (Ngb) and Hypoxia-inducible factor (HIF-1α) expression in the telencephalon of yak and cattle, and compare the expression and distribution pattern of Ngb and HIF-1α in the two animals. Immunohistochemistry (IHC), quantitative real-time Polymerase Chain Reaction (qRT-PCR), and Western blot (WB) were employed to investigate Ngb and Hif-1α expression in the telencephalon of yak and cattle. mRNA and protein expressions of Ngb and HIF-1α showed positive in different tissues of the yak and cattle telencephalon. Ngb expression in tissues of the yak recorded higher as compare to cattle while HIF-1α expression was found higher in cattle than yak. The HIF-1α expression in some tissues of yak telencephalon was consistent with the cattle. The results documented that HIF-1α may have a direct or indirect synergistic effect on Ngb expression in the yak telencephalon to improve hypoxia adaptation. It is suggested that yak may need more Ngb expression for adaptation, but the expression of HIF-1α seems to be down-regulated during long-term adaptation, and the specific causes of this phenomenon needs to be further verified.
O telencéfalo refere-se à parte anterior e mais desenvolvida do prosencéfalo, consistindo principalmente dos hemisférios cerebrais. O estudo determinou a expressão de neuroglobina (Ngb) e fator indutível por hipóxia (HIF-1α) no telencéfalo de iaques e bovinos e comparou a expressão e o padrão de distribuição de Ngb e HIF-1α nos dois animais. Imuno-histoquímica (IHC), reação em cadeia da polimerase quantitativa em tempo real (qRT-PCR) e Western blot (WB) foram empregados para investigar a expressão de Ngb e Hif-1α no telencéfalo de iaques e bovinos. As expressões de mRNA e proteínas de Ngb e HIF-1α mostraram-se positivas em diferentes tecidos do telencéfalo de iaque e bovino. A expressão de Ngb nos tecidos do iaque foi registrada mais alta em comparação com o gado, enquanto a expressão do HIF-1α foi encontrada mais alta no gado do que no iaque. A expressão de HIF-1α em alguns tecidos do telencéfalo de iaque foi consistente com o gado. Os resultados documentaram que o HIF-1α pode ter um efeito sinérgico direto ou indireto na expressão de Ngb no telencéfalo de iaque para melhorar a adaptação à hipóxia. É sugerido que o iaque pode precisar de mais expressão de Ngb para adaptação, mas a expressão de HIF-1α parece ser regulada para baixo durante a adaptação de longo prazo, e as causas específicas desse fenômeno precisam ser verificadas.
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Animales , Bovinos , Factor 1 Inducible por Hipoxia/análisis , Neuroglobina/análisis , Telencéfalo , Inmunohistoquímica , Reacción en Cadena en Tiempo Real de la Polimerasa , Western BlottingRESUMEN
Bioactive glasses (BGs) have shown great potential for tissue regeneration and their composition flexibility allows the incorporation of different ions with physiological activities and therapeutic properties in the glass network. Among the many ions that could be incorporated, cobalt (Co) is a significant one, as it mimics hypoxia, triggering the formation of new blood vessels by the vascular endothelial growth factor A (VEGFA), due to the stabilizing effect on the hypoxia inducible factor 1 subunit alpha (HIF1A), an activator of angiogenesis-related genes, and is therefore of great interest for tissue engineering applications. However, despite its promising properties, the effects of glasses incorporated with Co on angiogenesis, through human umbilical cord vein endothelial cells (HUVECs) studies, need to be further investigated. Therefore, this work aimed to evaluate the biocompatibility and angiogenic potential of a new sol-gel BG, derived from the SiO2 -CaO-P2 O5 -CoO system. The structural evaluation showed the predominance of an amorphous glass structure, and the homogeneous presence of cobalt in the samples was confirmed. in vitro experiments showed that Co-containing glasses did not affect the viability of HUVECs, stimulated the formation of tubes and the gene expression of HIF1A and VEGFA. in vivo experiments showed that Co-containing glasses stimulated VEGFA and HIF1A expression in blood vessels and cell nuclei, respectively, in the deep dermis layer of the dorsal region of rats, featuring considerable local stimulation of the angiogenesis process due to Co-release. Co-containing glasses showed therapeutic effect, and Co incorporation is a promising strategy for obtaining materials with superior angiogenesis properties for tissue engineering applications.
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Materiales Biomiméticos/química , Cobalto/química , Vidrio/química , Factor 1 Inducible por Hipoxia/análisis , Neovascularización Fisiológica , Factor A de Crecimiento Endotelial Vascular/análisis , Animales , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Materiales Biomiméticos/farmacología , Cobalto/farmacología , Células Endoteliales de la Vena Umbilical Humana , Humanos , Masculino , Neovascularización Fisiológica/efectos de los fármacos , Ratas WistarRESUMEN
OBJECTIVES: This study aimed to investigate the utility of fluorodeoxyglucose (FDG) positron emission tomography for solid pseudopapillary neoplasm (SPN) diagnosis. METHODS: The subjects included 53 cases of SPN. We compared the maximal standardized uptake volume (SUVmax) with those of 25 cases of pancreatic duct cancer and 18 cases of pancreatic neuroendocrine neoplasm. In addition, immunopathological testing for SPN with regard to FDG uptake was undertaken. RESULTS: An increase in SUVmax was observed in all tumors with increased tumor diameter. Among tumors of 20 mm or smaller, the SUVmax of SPN was significantly higher than those of pancreatic duct cancer and pancreatic neuroendocrine neoplasm. The results of a pathological study of FDG uptake in SPN revealed increased glucose transporter protein type 1 expression with tumor enlargement. Furthermore, increased hypoxia-inducible factor-1 and vascular endothelial growth factor expression under hypoxic conditions were observed in the areas of necrosis. CONCLUSIONS: In cases in which high FDG uptake is observed in small pancreatic tumors, FDG positron emission tomography is potentially useful for SPN differentiation. The factors involved in FDG uptake in SPN include cell density and glucose transporter protein expression, as well as hypoxia-inducible factor and vascular endothelia growth factor expression in the hypoxic environment of necrotic areas.
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Carcinoma Papilar/diagnóstico por imagen , Fluorodesoxiglucosa F18 , Neoplasias Pancreáticas/diagnóstico por imagen , Tomografía de Emisión de Positrones/métodos , Radiofármacos , Adolescente , Adulto , Anciano , Carcinoma Papilar/química , Carcinoma Papilar/patología , Niño , Femenino , Transportador de Glucosa de Tipo 1/análisis , Humanos , Factor 1 Inducible por Hipoxia/análisis , Masculino , Persona de Mediana Edad , Neoplasias Pancreáticas/química , Neoplasias Pancreáticas/patología , Factor A de Crecimiento Endotelial Vascular/análisis , Adulto Joven , Neoplasias PancreáticasRESUMEN
BACKGROUND: The present study was designed to investigate the effects of Berberis vulgaris (BV) juice consumption on plasma levels of insulin-like growth factor (IGF-1), IGF-binding proteins (IGFBPs), and the expression of PPAR-γ, VEGF and HIF in women with benign breast disease. METHODS: This parallel design randomized, double-blind controlled clinical trial was conducted on 85 eligible patients diagnosed with benign breast disease. They were assigned randomly into either BV juice group (n = 44, BV juice: 480 ml/day) or placebo group (n = 41, BV placebo juice: 480 ml/day) for 8 weeks intervention. Participants, caregivers and those who assessed laboratory analyses were blinded to the assignments. Plasma levels of biomarkers were measured at baseline and after 8 weeks by ELISA. Quantitative real-time PCR was used to measure the fold change in the expression of each interested gene. RESULTS: The compliance of participants was 95.2% and 40 available subjects analyzed in each group at last. Relative treatment (RT) effects for BV juice caused 16% fall in IGF-1 concentration and 37% reduction in the ratio of IGF-1/1GFBP1. Absolute treatment effect expressed 111 ng/ml increased mean differences of IGFBP-3 between BV group and placebo. Plasma level of PPAR-γ increased in both groups but it was not significant. Fold changes in the expressions of PPAR-γ, VEGF and HIF showed down-regulation in the intervention group compared to placebos (P < 0.05). CONCLUSIONS: The BV juice intervention over 8 weeks was accompanied by acceptable efficacy and decreased plasma IGF-1, and IGF-1/IGFBP-1 ratio partly could be assigned to enhanced IGFBP-1 level in women with BBD. The intervention caused reductions in the expression levels of PPAR, VEGF, and HIF which are remarkable genomic changes to potentially prevent breast tumorigenesis. TRIAL REGISTRATION: IRCT2012110511335N2. Registered 10 July 2013 (retrospectively registered).
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Berberis , Neoplasias de la Mama , Jugos de Frutas y Vegetales , Adulto , Mama/química , Mama/patología , Neoplasias de la Mama/química , Neoplasias de la Mama/dietoterapia , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Femenino , Humanos , Factor 1 Inducible por Hipoxia/análisis , Factor 1 Inducible por Hipoxia/metabolismo , Factor I del Crecimiento Similar a la Insulina/análisis , Factor I del Crecimiento Similar a la Insulina/metabolismo , Persona de Mediana Edad , PPAR gamma/análisis , PPAR gamma/metabolismo , Factor A de Crecimiento Endotelial Vascular/análisis , Factor A de Crecimiento Endotelial Vascular/metabolismo , Adulto JovenRESUMEN
Tuftelin was originally discovered and mostly studied in the tooth, but later found also in other organs. Despite its wide distribution among tissues, tuftelin's function has so far been specified only in the formation of enamel crystals. Nevertheless, in many cases, tuftelin was suggested to be associated with cellular adaptation to hypoxia and recently even with cell differentiation. Therefore, we aimed to investigate tuftelin expression along with hypoxia-inducible factors (HIFs) during the early development of the mandibular/alveolar (m/a) bone, when osteoblasts started to differentiate in vivo and to compare their expression levels in undifferentiated versus differentiated osteoblastic cells in vitro. Immunohistochemistry demonstrated the presence of tuftelin already in osteoblastic precursors which were also HIF1-positive, but HIF2-negative. Nevertheless, HIF2 protein appeared when osteoblasts differentiated, one day later. This is in agreement with observations made with MC3T3-E1 cells, where there was no significant difference in tuftelin and Hif1 expression in undifferentiated vs. differentiated cells, although Hif2 increased upon differentiation induction. In differentiated osteoblasts of the m/a bone, all three proteins accumulated, first, prenatally, in the cytoplasm and later, particularly at postnatal stages, they displayed also peri/nuclear localization. Such a dynamic time-space pattern of tuftelin expression has recently been reported in neurons, which, as the m/a bone, differentiate under less hypoxic conditions as indicated also by a prevalent cytoplasmic expression of HIF1 in osteoblasts. However, unlike what was shown in cultured neurons, tuftelin does not seem to participate in final osteoblastic differentiation and its functions, thus, appears to be tissue specific.
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Proteínas del Esmalte Dental/análisis , Factor 1 Inducible por Hipoxia/análisis , Osteogénesis/genética , Factores de Transcripción/análisis , Células 3T3 , Animales , Células Cultivadas , Proteínas del Esmalte Dental/genética , Factor 1 Inducible por Hipoxia/genética , Inmunohistoquímica , Ratones , Factores de Transcripción/genéticaRESUMEN
BACKGROUND The timing of parturition is an important determinant of labor and delivery care. Early parturition is associated with increased neonatal morbidity and mortality. Most existing studies analyzed a single factor for the initiation of parturition, and the role of multiple factors in initiating parturition has not been comprehensively analyzed. MATERIAL AND METHODS We measured the levels of proinflammatory mediators, hypoxia factor, matrix metalloproteinases, hormones, and oxytocin, as well as fetal umbilical blood flow, before and after labor, and their associations with parturition. We also built a statistical model to predict the timing of parturition based on the measurement data. RESULTS IL-1ß, IL-6, TNF-alpha, MMP-9, and HIF-1alpha concentrations significantly increased from full term to labor. The PRL level significantly decreased from full term to parturition. There was no significant change in MCP-1, E3, and OT concentrations from full term to parturition. IL-1ß, IL-6, TNF-alpha, and MMP-9 concentrations were negatively correlated with the initiation of parturition. There was a small but nonsignificant increase in umbilical venous blood flow before parturition. Multiple factors showed a close correlation with the initiation of parturition, and area under the curve analysis showed that a multiple factor model was superior to single factors in the establishment of a model to predict initiation of parturition; however, these results need further confirmation. CONCLUSIONS Combined proinflammatory biomarkers have better predictive value for term labor than single biomarkers.
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Predicción/métodos , Parto/metabolismo , Nacimiento a Término/metabolismo , Biomarcadores/sangre , Parto Obstétrico , Femenino , Sangre Fetal , Edad Gestacional , Hormonas/análisis , Hormonas/metabolismo , Humanos , Factor 1 Inducible por Hipoxia/análisis , Factor 1 Inducible por Hipoxia/metabolismo , Inflamación/metabolismo , Trabajo de Parto , Metaloproteinasas de la Matriz/análisis , Metaloproteinasas de la Matriz/metabolismo , Modelos Estadísticos , Oxitocina/análisis , Oxitocina/metabolismo , Parto/fisiología , Embarazo , Nacimiento a Término/fisiologíaRESUMEN
The authors describe a significantly improved colorimetric nanoprobe for the determination of transcription factors (TFs). It is making use of click-mediated growth of gold nanoparticles (AuNPs) to amplify the signal-to-noise ratio. Hypoxia-inducible factor-1 (HIF-1) is an important TF that acts as a mediator of cell response to hypoxia. So, the detection of HIF-1 was chosen as the model analyte. Specifically, target HIF-1 is designed to bind to the hypoxia response element within DNA duplex. The click chemistry between the DNA duplex and alkynyl-functionalized AuNPs (AF-AuNPs) is then inhibited because of significant steric hindrance. As a result, the AF-AuNPs grow into larger-sized highly-aggregated irregular nanostructures, which in turn enable colorimetric determination. The ratio of absorbances at 620 and 560 nm increases in the 0.5 to 10 nM HIF-1 concentration range, and the detection limit is 0.27 nM. This is better by a factor of 100 than that of aggregation-based colorimetric assays. The nanoprobe is selective and can be used in complex samples. Conceivably, it may also be extended to the determination of other TFs by simply changing the used DNA duplex. Graphical abstract Schematic of a nanoprobe for detecting hypoxia-inducible factor-1 (HIF-1). Three concepts are involved: the binding of HIF-1 and hypoxia response element, the Cu+-catalyzed click chemistry between P1/P2 duplex and alkynyl-functionalized AuNPs (AF-AuNPs), and the AuNPs growth with hydroxylamine and HAuCl4.
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Colorimetría/métodos , Oro/química , Factor 1 Inducible por Hipoxia/análisis , Nanopartículas del Metal/química , Química Clic , Humanos , Límite de DetecciónAsunto(s)
Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Factor 1 Inducible por Hipoxia/metabolismo , Sepsis/complicaciones , Trombosis/etiología , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/análisis , Hipoxia de la Célula , Humanos , Factor 1 Inducible por Hipoxia/análisis , Sepsis/metabolismo , Transducción de Señal , Trombosis/metabolismoRESUMEN
OBJECTIVE: Grape seed proanthocyanidine extract (GSPE) is a strong antioxidant derived from the grape seeds (Vitis vinifera, Terral J.F.) and has a polyphenolic structure with a wide range of biological activity. The aim of the present study was to evaluate the effects of GSPE on alveolar bone loss and histopathological changes in rats with diabetes mellitus and ligature-induced periodontitis. MATERIAL AND METHODS: Forty rats were divided into 6 study groups. Control (C, 6 rats) group, periodontitis (P, 6 rats) group, diabetes (D, 6 rats) group, diabetes and periodontitis (D+P, 6 rats) group, diabetes, periodontitis and 100 mg/kg/day GSPE (GSPE-100, 8 rats), and diabetes, periodontitis and 200 mg/kg/day GSPE (GSPE-200, 8 rats) group. Diabetes mellitus was induced by intraperitoneal injection of a single dose of streptozotocin (60 mg/kg). Periodontitis was induced via ligation method. Silk ligatures were placed at the mandibular right first molars. GSPE was administered by oral gavage. After 30 days, all rats were killed. Alveolar bone loss was measured morphometrically via a stereomicroscope. For histopathological analyses, Alizarin red staining, and matrix metalloproteinase (MMP)-8, vascular endothelial growth factor and hypoxia inducible factor (HIF)-1α immunohistochemistry were performed. Tartrate-resistant acid phosphatase-positive osteoclast cells and relative total inflammatory cells were also determined. RESULTS: The highest alveolar bone loss was observed in the D+P group (P < .05). GSP-200 group decreased alveolar bone loss (P < .05). The D+P group had the highest osteoclast counts, but the difference was not significant compared to the P, GSPE-100 and GSPE-200 groups (P > .05). The inflammation in the D+P group was also higher than the other groups (P < .05). The osteoblast numbers increased in the GSPE-100 and GSPE-200 groups compared to the P and D+P groups (P < .05). MMP-8 and HIF-1α levels were highest in the D+P group and GSPE significantly decreased these levels (P < .05). CONCLUSION: Within the limits of this animal study, it can be suggested that GSPE administration may decrease periodontal inflammation and alveolar bone loss via decreasing MMP-8 and HIF-1α levels and increase osteoblastic activity in diabetic rats with experimental periodontitis.
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Pérdida de Hueso Alveolar/tratamiento farmacológico , Pérdida de Hueso Alveolar/patología , Diabetes Mellitus Experimental/complicaciones , Extracto de Semillas de Uva/farmacología , Extracto de Semillas de Uva/uso terapéutico , Periodontitis/tratamiento farmacológico , Periodontitis/patología , Proantocianidinas/farmacología , Proantocianidinas/uso terapéutico , Pérdida de Hueso Alveolar/clasificación , Proceso Alveolar/patología , Animales , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Glucemia/análisis , Peso Corporal , Modelos Animales de Enfermedad , Extracto de Semillas de Uva/administración & dosificación , Factor 1 Inducible por Hipoxia/análisis , Inmunohistoquímica , Inflamación/tratamiento farmacológico , Inflamación/patología , Inyecciones Intraperitoneales , Ligadura/efectos adversos , Masculino , Metaloproteinasa 8 de la Matriz/análisis , Osteoblastos/efectos de los fármacos , Osteoblastos/patología , Osteoclastos/efectos de los fármacos , Osteoclastos/patología , Proantocianidinas/administración & dosificación , Ratas , Ratas Wistar , Estreptozocina/administración & dosificación , Estreptozocina/farmacología , Fosfatasa Ácida Tartratorresistente/análisis , Factor A de Crecimiento Endotelial Vascular/análisisRESUMEN
The complex cross-talk of intricate intercellular signaling networks between the tumor and stromal cells promotes cancer progression. Hypoxia is one of the most common conditions encountered within the tumor microenvironment that drives tumorigenesis. Most responses to hypoxia are elicited by a family of transcription factors called hypoxia-inducible factors (HIFs), which induce expression of a diverse set of genes that assist cells to adapt to hypoxic environments. Among the three HIF protein family members, the role of HIF-1 is well established in cancer progression. HIF-1 functions as a signaling hub to coordinate the activities of many transcription factors and signaling molecules that impact tumorigenesis. This mini review discusses the complex role of HIF-1 and its context-dependent partners under various cancer-promoting events including inflammation and generation of cancer stem cells, which are implicated in tumor metastasis and relapse. In addition, the review highlights the importance of therapeutic targeting of HIF-1 for cancer prevention.
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Hipoxia de la Célula , Factor 1 Inducible por Hipoxia/fisiología , Inflamación/etiología , Neoplasias/etiología , Animales , Transición Epitelial-Mesenquimal , Humanos , Factor 1 Inducible por Hipoxia/análisis , Células Madre Neoplásicas/fisiología , Especies Reactivas de Oxígeno/metabolismo , Microambiente TumoralRESUMEN
BACKGROUND: Although the function of hypoxia-inducible factor 1 (HIF1) in many kinds of solid tumor has been revealed, the significance of HIF1 in osteosarcoma is still controversial and not well understood. METHODS: Immunohistochemistry was used to detect HIF1 expression. The correlation between HIF1 and clinicopathology factors was analyzed by use of chi-squared tests. The prognostic value of HIF1 was evaluated by univariate and multivariate analysis. Moreover, the function of HIF1 in osteosarcoma cells was further investigated in in-vitro experiments by regulating HIF1 and vascular endothelial growth factor-A (VEGF-A) expression. RESULTS: Expression of HIF1 was high for 56.82 % of the samples in our investigation. HIF1 expression was significantly associated with positive metastasis (P = 0.037). By use of the Kaplan-Meier method, high expression of HIF1 was proved to be related to poorer overall survival (P = 0.007). By use of a Cox-regression model, HIF1 was identified as an independent prognostic biomarker (P = 0.019). We also proved that HIF1 can promote osteosarcoma invasion in hypoxia by inducing VEGF-A expression. CONCLUSIONS: HIF1 was identified as an independent prognostic biomarker in osteosarcoma. It can promote osteosarcoma cell invasion by inducing VEGF-A expression, indicating that HIF1 is a potential drug target in osteosarcoma.
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Neoplasias Óseas/química , Factor 1 Inducible por Hipoxia/análisis , Osteosarcoma/química , Osteosarcoma/secundario , Factor A de Crecimiento Endotelial Vascular/análisis , Biomarcadores de Tumor/análisis , Neoplasias Óseas/patología , Hipoxia de la Célula , Línea Celular Tumoral , Femenino , Técnicas de Silenciamiento del Gen , Humanos , Factor 1 Inducible por Hipoxia/genética , Factor 1 Inducible por Hipoxia/metabolismo , Inmunohistoquímica , Estimación de Kaplan-Meier , Masculino , Invasividad Neoplásica , Pronóstico , Modelos de Riesgos Proporcionales , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismo , Adulto JovenRESUMEN
BACKGROUND: Angiogenesis, which is initiated by certain tumor micro-environmental conditions and diverse protein factors, plays a pivotal role during tumor development and metastasis. Therefore, many efforts have been made to develop effective anti-angiogenic agents as anticancer therapeutics. In the current study, we investigated the anti-angiogenic potential of an ethanol extract of Annona atemoya seeds (EEAA) in vitro and in vivo. METHODS: The anti-angiogenic potential of EEAA was evaluated using various in vitro/in vivo models, including cell proliferation, migration, and tube formation by human umbilical vascular endothelial cells (HUVECs); a Matrigel plug assay; and tumor-induced angiogenesis. The expression of hypoxia-inducible factors (HIFs) and vascular endothelial growth factor (VEGF) was investigated using reverse transcription-polymerase chain reaction, immunoassays, and western blotting. RESULTS: EEAA was able to significantly inhibit the angiogenic properties of HUVECs in vitro as well as angiogenic factor-induced blood vessel formation in vivo. EEAA down-regulated the expression of VEGF and HIF-1alpha/2alpha at the mRNA and protein levels, respectively, in cancer cells under hypoxic conditions. CONCLUSIONS: EEAA shows a strong anti-angiogenic potential in both in vitro and in vivo systems, and we suggest that EEAA may be a valuable herbal source for anticancer drug development.
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Inhibidores de la Angiogénesis/farmacología , Annona/química , Extractos Vegetales/farmacología , Semillas/química , Análisis de Varianza , Inhibidores de la Angiogénesis/química , Animales , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Etanol , Femenino , Células Endoteliales de la Vena Umbilical Humana , Humanos , Factor 1 Inducible por Hipoxia/análisis , Factor 1 Inducible por Hipoxia/genética , Factor 1 Inducible por Hipoxia/metabolismo , Ratones , Ratones Desnudos , Extractos Vegetales/química , Factor A de Crecimiento Endotelial Vascular/análisis , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
BACKGROUND: The skin color of human hypertrophic scar changes dynamically during scar progression. OBJECTIVE: To investigate whether hypoxia is dynamic during scar progression. METHODS: Thirty-five patients with early, proliferative, regressive, and mature scars were involved in this study. Tissue oxygen tension was measured before scar surgery. After surgery, the scar stage was further defined using hematoxylin and eosin staining, and microvessel density and hypoxia inducible factor-1 (HIF-1) expression were detected using immunohistochemistry to determine a correlation with oxygen level. RESULTS: Mild hypoxia is present in early scars, moderate hypoxia in proliferative scars, and severe hypoxia in regressive scars. Oxygen levels then return to normal in mature scars, which was consistent with the dynamic change in microvessel density. Meanwhile, HIF-1 expression also changed dynamically along with alteration in oxygen levels. CONCLUSION: Hypoxia is dynamic in scar tissue and is possibly correlated with scar formation and regression.
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Cicatriz Hipertrófica/sangre , Cicatriz Hipertrófica/patología , Progresión de la Enfermedad , Oxígeno/metabolismo , Adolescente , Adulto , Antígenos CD34/análisis , Monitoreo de Gas Sanguíneo Transcutáneo , Hipoxia de la Célula , Colorantes , Eosina Amarillenta-(YS) , Femenino , Hematoxilina , Humanos , Factor 1 Inducible por Hipoxia/análisis , Masculino , Microvasos , Persona de Mediana Edad , Coloración y Etiquetado , Adulto JovenRESUMEN
INTRODUCTION: Human dental pulp cells (HDPCs) are recalcitrant to hypoxic stress. We investigated whether hypoxia-induced autophagy of HDPCs offered these cells a survival advantage and the underlying mechanism of this resistance. METHODS: The viability and apoptosis of HDPCs were examined after exposure to hypoxia by Vi-CELL cell viability analyzer and flow cytometry. Autophagy was assessed by using immunofluorescence, acridine orange staining, real-time polymerase chain reaction, and Western blotting. Either 3-methyladenine or expression vectors encoding dominant negative ATG5 were used to inhibit autophagy. Rapamycin was used as an autophagic inducer. To explore the mechanisms of autophagy, adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway and hypoxia-inducible transcription factor-1 were suppressed by chemical inhibitors Compound C and YC-1, respectively. RESULTS: The exposure of HDPCs to hypoxia had no effect on viability and resulted in increasing acidic vesicular organelle-positive cells, autophagosome formation, and up-regulation of autophagy genes. Inhibition of autophagy with 3- methyladenine or expression vectors encoding dominant negative ATG5 abrogated the protective effects of HDPCs. The phosphorylation of AMPK was up-regulated, whereas the phosphorylation of mTOR was down-regulated in hypoxia-treated HDPCs, which were both attenuated by Compound C. Furthermore, treatment with Compound C rather than YC-1 reduced the autophagy. CONCLUSIONS: Our results suggested that autophagy of HDPCs might be cytoprotective against hypoxic stress via the AMPK/mTOR signaling pathway.
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Proteínas Quinasas Activadas por AMP/fisiología , Autofagia/fisiología , Hipoxia de la Célula/fisiología , Pulpa Dental/enzimología , Serina-Treonina Quinasas TOR/fisiología , Proteínas Quinasas Activadas por AMP/antagonistas & inhibidores , Adenina/análogos & derivados , Adenina/farmacología , Apoptosis/fisiología , Proteínas Reguladoras de la Apoptosis/análisis , Autofagia/efectos de los fármacos , Proteína 12 Relacionada con la Autofagia , Proteína 5 Relacionada con la Autofagia , Beclina-1 , Técnicas de Cultivo de Célula , Supervivencia Celular/fisiología , Células Cultivadas , Pulpa Dental/citología , Vectores Genéticos/genética , Guanilato Ciclasa/farmacología , Humanos , Factor 1 Inducible por Hipoxia/análisis , Factor 1 Inducible por Hipoxia/antagonistas & inhibidores , Indazoles/farmacología , Proteínas de la Membrana/análisis , Proteínas Asociadas a Microtúbulos/análisis , Plásmidos/genética , Proteínas Proto-Oncogénicas/análisis , Proteínas Modificadoras Pequeñas Relacionadas con Ubiquitina/análisis , Serina-Treonina Quinasas TOR/antagonistas & inhibidores , Proteínas Supresoras de Tumor/análisisRESUMEN
PURPOSE: We aimed to develop a radiolabeled peptide probe for the imaging of hypoxia-inducible factor-1 (HIF-1)-active tumors. PROCEDURES: We synthesized the peptide probes that contain or lack an essential sequence of the oxygen-dependent degradation of HIF-1α in proteasomes ((123/125)I-DKOP30 or (125)I-mDKOP, respectively). The degradation of probes was evaluated in vitro using cell lysates containing proteasomes. In vivo biodistribution study, planar imaging, autoradiography, and comparison between probe accumulation and HIF-1 transcriptional activity were also performed. RESULTS: The (125)I-DKOP30 underwent degradation in a proteasome-dependent manner, while (125)I-mDKOP was not degraded. Biodistribution analysis showed (125)I-DKOP30 accumulation in tumors. The tumors were clearly visualized by in vivo imaging, and intratumoral distribution of (125)I-DKOP30 coincided with the HIF-1α-positive hypoxic regions. Tumoral accumulation of (125)I-DKOP30 was significantly correlated with HIF-1-dependent luciferase bioluminescence, while that of (125)I-mDKOP was not. CONCLUSION: (123)I-DKOP30 is a useful peptide probe for the imaging of HIF-1-active tumors.
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Factor 1 Inducible por Hipoxia/análisis , Factor 1 Inducible por Hipoxia/metabolismo , Técnicas de Sonda Molecular , Sondas Moleculares/farmacocinética , Péptidos/química , Secuencia de Aminoácidos , Animales , Autorradiografía , Hipoxia de la Célula , Línea Celular Tumoral , Femenino , Células HeLa , Humanos , Factor 1 Inducible por Hipoxia/química , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones Desnudos , Sondas Moleculares/química , Datos de Secuencia Molecular , Péptidos/farmacocinética , Distribución Tisular , Tomografía Computarizada de Emisión de Fotón ÚnicoRESUMEN
PURPOSE: Oral pregnancy tumors (OPTs) arise on the inflamed gingiva of women after the first trimester of pregnancy. The expression of angiogenic markers and female hormone receptors was assessed. MATERIALS AND METHODS: Immunohistochemistry was used to analyze the expression of estrogen and progesterone receptors and the expression of angiogenic factors, such as vascular endothelial growth factor (VEGF) and its receptor, fibroblast growth factor (FGF), and hypoxia inducible factors 1α and 3α (HIF1α and HIF3α). Experimental groups included 9 OPTs, 10 oral pyogenic granulomas from nonpregnant women of the same age, and 9 oral pyogenic granulomas from postmenopausal women. RESULTS: VEGF expression in stromal histiocytes and endothelial cells of small vessels was positively correlated in the OPT group (P < .05 by χ(2) test). VEGF receptor also was overexpressed in stromal histiocytes and endothelial cells of OPTs compared with oral pyogenic granulomas from nonpregnant and postmenopausal women (P < .005 by χ(2) test). No correlation was detected among estrogen and progesterone receptors, FGF and HIF1α and HIF3α (ER and PgR respectively) in the 3 experimental groups. CONCLUSIONS: VEGF-associated angiogenesis is most likely involved in the pathogenesis of the lesion. These results imply that local inhibition of VEGF activity could be an adjuvant therapeutic approach for OPTs to control hemorrhage, which can be massive at the surgical excision of such lesions during pregnancy.
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Inductores de la Angiogénesis/análisis , Neoplasias Gingivales/metabolismo , Factor 1 Inducible por Hipoxia/análisis , Neovascularización Patológica/metabolismo , Complicaciones Neoplásicas del Embarazo/metabolismo , Receptores de Progesterona/biosíntesis , Receptores de Factores de Crecimiento Endotelial Vascular/biosíntesis , Factor A de Crecimiento Endotelial Vascular/biosíntesis , Adulto , Estudios de Casos y Controles , Distribución de Chi-Cuadrado , Femenino , Factores de Crecimiento de Fibroblastos/análisis , Neoplasias Gingivales/complicaciones , Granuloma Piogénico/metabolismo , Humanos , Persona de Mediana Edad , Neovascularización Patológica/complicaciones , Posmenopausia , Embarazo , Receptores de Estrógenos/biosíntesis , Adulto JovenRESUMEN
Diaromatic-substituted ortho- and meta-carboranes were synthesized as mimics of manassantin A. Among the carboranes synthesized, compounds 1 and 2 showed significant inhibition of hypoxia-induced HIF-1 transcriptional activity, with IC(50) values of 3.2 and 2.2 µM, respectively. Compounds 1 and 2 similarly suppressed hypoxia-induced HIF-1α accumulation in a concentration-dependent manner without affecting the expression level of HIF-1α mRNA. The hypoxia-induced accumulation and translocation of HIF-1α into nuclei were not observed in HeLa cells treated with compounds 1 and 2 by immunofluorescence analysis, revealing that the inhibition of hypoxia-induced HIF-1 transcriptional activity is induced by compounds 1 and 2 through a degradation pathway of the HIF-1α protein under hypoxic conditions.
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Compuestos de Boro/química , Compuestos de Boro/farmacología , Factor 1 Inducible por Hipoxia/genética , Lignanos/química , Lignanos/farmacología , Activación Transcripcional/efectos de los fármacos , Hipoxia de la Célula/efectos de los fármacos , Línea Celular Tumoral , Células HeLa , Humanos , Factor 1 Inducible por Hipoxia/análisis , Factor 1 Inducible por Hipoxia/metabolismo , ARN Mensajero/genética , Transducción de Señal/efectos de los fármacosRESUMEN
BACKGROUND: Spermatic cord torsion can lead to testis ischemia (I) and subsequent ischemia-reperfusion (I/R) causing germ cell-specific apoptosis. Previously, we demonstrated that the hypoxia-inducible factor-1 (HIF-1) transcription factor, a key regulator of physiological responses to hypoxia, is abundant in Leydig cells in normoxic and ischemic testes. We hypothesize that testicular HIF-1 activates the expression of antiapoptotic target genes to protect Leydig cells from apoptosis. In silico analysis of testis genes containing a consensus hypoxia response element (HRE, 5'-RCGTG-3') identified myeloid cell leukemia-1 (Mcl-1) as a potential HIF-1 target gene. The purpose of this study was to determine whether HIF-1 shows DNA-binding activity in normoxic and ischemic testes and whether Mcl-1 is a target gene of testicular HIF-1. METHODS: The testicular HIF-1 DNA-binding capacity was analyzed in vitro using a quantitative enzyme-linked immunosorbent assay (ELISA) and electrophoretic mobility shift assays (EMSA). MCL-1 protein expression was evaluated by immunoblot analysis and immunohistochemistry. The binding of testicular HIF-1 to the Mcl-1 gene was examined via chromatin immunoprecipitation (ChIP) analysis. RESULTS: The ELISA and EMSA assays demonstrated that testicular HIF-1 from normoxic and ischemic testes binds DNA equally strongly, suggesting physiological roles for HIF-1 in the normoxic testis, unlike most tissues in which HIF-1 is degraded under normoxic conditions and is only activated by hypoxia. MCL-1 protein was determined to be abundant in both normoxic and ischemic testes and expressed in Leydig cells. In a pattern identical to that of HIF-1 expression, the steady-state levels of MCL-1 were not significantly affected by I or I/R and MCL-1 co-localized with HIF-1α in Leydig cells. Chromatin immunoprecipitation (ChIP) analysis using a HIF-1 antibody revealed sequences enriched for the Mcl-1 promoter. CONCLUSIONS: The results demonstrated that, unlike what is observed in most tissues, HIF-1 displays DNA-binding activity in both normoxic and ischemic testes, and Mcl-1 may be a key target gene of testicular HIF-1 with potential roles in the antiapoptotic protection of Leydig cells.
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Factor 1 Inducible por Hipoxia/análisis , Factor 1 Inducible por Hipoxia/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/genética , Testículo/química , Animales , Apoptosis , Hipoxia de la Célula , Inmunoprecipitación de Cromatina , ADN/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/análisis , Immunoblotting , Inmunohistoquímica , Isquemia/metabolismo , Células Intersticiales del Testículo/química , Masculino , Proteína 1 de la Secuencia de Leucemia de Células Mieloides , Proteínas Proto-Oncogénicas c-bcl-2/análisis , Ratas , Ratas Sprague-Dawley , Testículo/irrigación sanguíneaRESUMEN
The purpose of this paper is to explore neovascularization in the biomaterials implanted in vivo. Capillaries formation of dermis defect filled with porous silk fibroin films (PSFFs) implant were investigated in rat at different times. Vascular endothelial growth factor (VEGF), CD34 and hypoxia inducible factor-1 (HIF) were observed in inflammatory cells and fibroblast in PSFFs by day 1, but expression in HIF and VEGF decreased by day 7, otherwise VEGF expressed in vascular endothelial cell increased by day 15 and 25, respectively. VEGF, angiopoietin 1 (Ang-1) and integrins αv and integrins ß3 in PSFFs were observed by Reverse transcription-polymerase chain reaction (RT-PCR). Polyvinyl alcohol (PVA) sponge was control. PSFFs have better neovascularization than that of PVA expression of VEGF, HIF and angiogenesis in immunohistochemical and mRNA level.