Abstract
Aims:
Our aims were to determine the role of Notch1 in mediating visfatin-induced angiogenesis and to explore potential target genes involved.
Methods and results:
Inhibition of Notch signalling attenuated visfatin-induced angiogenesis in vitro, ex vivo, and in vivo. Visfatin increased γ-secretase activity, Notch1 cleavage and activation, and Hes1 gene induction. Visfatin also stimulated fibroblast growth factor-2 (FGF-2) gene expression in a Notch1-dependent manner. Enforced expression of active Notch1 intracellular domain increased FGF-2 protein levels and stimulated endothelial tube formation, whereas blocking Notch1 signalling or knockdown of Notch1 by small interfering RNA suppressed visfatin-induced FGF-2 up-regulation and angiogenesis. Reporter analysis of FGF-2 promoter revealed the presence of CSL (CBF-1, suppressor of hairless, LAG-1)-binding site, and chromatin immunoprecipitation analysis demonstrated the binding of Notch1-CSL complex to this site in response to visfatin.
Conclusion:
Our data provide the first example of Notch1-dependent endothelial FGF-2 induction by visfatin and of Notch1 activation in visfatin-stimulated endothelial angiogenesis, suggesting that the signalling axis of visfatin/Notch1/angiogenic factors like FGF-2 might be a valuable target for pathological angiogenesis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amyloid Precursor Protein Secretases / antagonists & inhibitors
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Amyloid Precursor Protein Secretases / metabolism
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Animals
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Basic Helix-Loop-Helix Transcription Factors / genetics
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Binding Sites
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Cells, Cultured
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Chick Embryo
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Chromatin Immunoprecipitation
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Cytokines / metabolism*
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Dipeptides / pharmacology
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Endothelial Cells / drug effects
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Endothelial Cells / enzymology*
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Enzyme Inhibitors / pharmacology
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Fibroblast Growth Factor 2 / genetics
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Fibroblast Growth Factor 2 / metabolism*
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Genes, Reporter
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Homeodomain Proteins / genetics
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Humans
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Immunoglobulin J Recombination Signal Sequence-Binding Protein / metabolism
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Male
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Mice
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Mice, Inbred C57BL
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Neovascularization, Physiologic* / drug effects
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Nicotinamide Phosphoribosyltransferase / metabolism*
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Promoter Regions, Genetic
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RNA Interference
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Rats
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Rats, Sprague-Dawley
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Receptor, Notch1 / genetics
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Receptor, Notch1 / metabolism*
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Recombinant Proteins / metabolism
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Signal Transduction
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Transcription Factor HES-1
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Transfection
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Up-Regulation
Substances
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Basic Helix-Loop-Helix Transcription Factors
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Cytokines
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Dipeptides
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Enzyme Inhibitors
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Homeodomain Proteins
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Immunoglobulin J Recombination Signal Sequence-Binding Protein
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N-(N-(3,5-difluorophenacetyl)alanyl)phenylglycine tert-butyl ester
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NOTCH1 protein, human
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RBPJ protein, human
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Receptor, Notch1
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Recombinant Proteins
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Transcription Factor HES-1
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Fibroblast Growth Factor 2
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HES1 protein, human
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Nicotinamide Phosphoribosyltransferase
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nicotinamide phosphoribosyltransferase, human
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Amyloid Precursor Protein Secretases