Activation of vascular endothelial growth factor through reactive oxygen species mediates 20-hydroxyeicosatetraenoic acid-induced endothelial cell proliferation

Austin M. Guo, Ali Syed Arbab, John R. Falck, Ping Chen, Paul A. Edwards, Richard J. Roman, A. Guillermo Scicli

Research output: Contribution to journalArticle

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Abstract

20-Hydroxyeicosatetraenoic acid (20-HETE) is formed by the ω-hydroxylation of arachidonic acid by cytochrome P450 4A and 4F enzymes, and it induces angiogenic responses in vivo. To test the hypothesis that 20-HETE increases endothelial cell (EC) proliferation via vascular endothelial growth factor (VEGF), we studied the effects of WIT003 [20-hydroxyeicosa-5(Z),14(Z)- dienoic acid], a 20-HETE analog on human macrovascular or microvascular EC. WIT003, as well as pure 20-HETE, stimulated EC proliferation by ∼40%. These proliferative effects were accompanied by increased VEGF expression and release that were observed as early as 4 h after 20-HETE agonist addition. This was accompanied by increased phosphorylation of the VEGF receptor 2. The proliferative effects of 20-HETE were markedly inhibited by a VEGF-neutralizing antibody. Polyethylene glycol-superoxide dismutase (PEG-SOD) markedly inhibited both the increases in VEGF expression and the proliferative effects of 20-HETE. In contrast, administration of the NAD(P)H oxidase inhibitor apocynin had no effect to the proliferative response to 20-HETE. The 20-HETE agonist markedly increased superoxide formation as reflected by an increase in dihydroethidium staining of EC, and this increase was inhibited by PEG-SOD but not by apocynin. 20-HETE also increased the phosphorylation of p42/p44 mitogen-activated protein kinase MAPK) in EC, whereas an inhibitor of MAPK [U0126, 1,4-diamino-2,3- dicyano-1,4-bis(2-aminophenylthio)butadiene] suppressed the proliferative and the VEGF changes but not the pro-oxidant effects of 20-HETE. These data suggest that 20-HETE stimulates superoxide formation by pathways other than apocynin-sensitive NAD(P)H oxidase, thereby activating MAPK and then enhancing VEGF synthesis that drives EC proliferation. Thus, 20-HETE may be involved in the regulation of EC functions, such as angiogenesis.

Original languageEnglish (US)
Pages (from-to)18-27
Number of pages10
JournalJournal of Pharmacology and Experimental Therapeutics
Volume321
Issue number1
DOIs
StatePublished - Apr 1 2007
Externally publishedYes

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Vascular Endothelial Growth Factor A
Reactive Oxygen Species
Endothelial Cells
Cell Proliferation
NADPH Oxidase
Superoxides
20-hydroxy-5,8,11,14-eicosatetraenoic acid
Phosphorylation
Vascular Endothelial Growth Factor Receptor-2
Mitogen-Activated Protein Kinase 1
Hydroxylation
Neutralizing Antibodies
Arachidonic Acid
Cytochrome P-450 Enzyme System
Staining and Labeling

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

Cite this

Activation of vascular endothelial growth factor through reactive oxygen species mediates 20-hydroxyeicosatetraenoic acid-induced endothelial cell proliferation. / Guo, Austin M.; Arbab, Ali Syed; Falck, John R.; Chen, Ping; Edwards, Paul A.; Roman, Richard J.; Scicli, A. Guillermo.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 321, No. 1, 01.04.2007, p. 18-27.

Research output: Contribution to journalArticle

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AU - Falck, John R.

AU - Chen, Ping

AU - Edwards, Paul A.

AU - Roman, Richard J.

AU - Scicli, A. Guillermo

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