In vivo expression of recombinant vascular endothelial growth factor in rabbit carotid artery increases production of superoxide anion

Arterioscler Thromb Vasc Biol. 2005 Mar;25(3):506-11. doi: 10.1161/01.ATV.0000153284.81572.f0. Epub 2004 Dec 16.

Abstract

Objective: Vascular endothelial growth factor (VEGF) is one of the most important pro-angiogenic cytokines. Ability of VEGF to stimulate formation of superoxide anion in vivo has not been studied. We hypothesized that in vivo expression of recombinant VEGF in the rabbit carotid artery increases production of superoxide anion.

Methods and results: Plaque-forming units (10(9)) of adenovirus-encoding human VEGF165 (AdVEGF) or beta-galactosidase (AdLacZ) were delivered into the lumen of rabbit carotid arteries. Three days after gene delivery, expression of recombinant proteins was detected in endothelium and smooth muscle cells. Endothelium-dependent relaxations to acetylcholine were impaired in AdVEGF-transduced arteries (P<0.01; n=5). Treatment with superoxide dismutase mimetic, Mn(III) tetra(4-benzoic acid) porphyrin chloride (10(-5) mol/L), improved relaxations to acetylcholine (P<0.01; n=5). Western blot analysis demonstrated increased expression of p47(phox) in AdVEGF-transduced arteries (P<0.05; n=8). Lucigenin chemiluminescence showed significantly higher production of superoxide anion in AdVEGF-transduced arteries (P<0.05; n=5 to 10).

Conclusions: Our results suggest that in vivo expression of recombinant VEGF in the vascular endothelium increases local production of superoxide anion. Superoxide anion appears to be an important mediator of vascular effects of VEGF in vivo.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Biopterins / genetics
  • Carotid Arteries / cytology
  • Carotid Arteries / metabolism*
  • Carotid Arteries / physiology*
  • Cyclic GMP / metabolism
  • GTP Cyclohydrolase / metabolism
  • Gene Transfer Techniques
  • Humans
  • Hydrazines / pharmacology
  • Immunohistochemistry
  • Male
  • NADPH Oxidases / metabolism
  • Nitric Oxide Donors / pharmacology
  • Nitric Oxide Synthase / metabolism
  • Nitric Oxide Synthase Type III
  • Nitrogen Oxides
  • Phenylephrine / pharmacology
  • Phosphoproteins / metabolism
  • Rabbits
  • Recombinant Proteins / genetics
  • Superoxides / metabolism*
  • Vascular Endothelial Growth Factor A / genetics*
  • Vascular Endothelial Growth Factor A / metabolism
  • Vasoconstriction / drug effects
  • Vasoconstriction / physiology
  • Vasoconstrictor Agents / pharmacology

Substances

  • Hydrazines
  • Nitric Oxide Donors
  • Nitrogen Oxides
  • Phosphoproteins
  • Recombinant Proteins
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • Vasoconstrictor Agents
  • Superoxides
  • Phenylephrine
  • Biopterins
  • 1,1-diethyl-2-hydroxy-2-nitrosohydrazine
  • NOS3 protein, human
  • Nitric Oxide Synthase
  • Nitric Oxide Synthase Type III
  • NADPH Oxidases
  • neutrophil cytosolic factor 1
  • GTP Cyclohydrolase
  • Cyclic GMP