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on December 16, 2004

Arteriosclerosis, Thrombosis, and Vascular Biology. 2004
Published online before print December 16, 2004, doi: 10.1161/01.ATV.0000153284.81572.f0
A more recent version of this article appeared on March 1, 2005
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Submitted on August 3, 2004
Accepted on November 22, 2004

In Vivo Expression of Recombinant Vascular Endothelial Growth Factor in Rabbit Carotid Artery Increases Production of Superoxide Anion

Osamu Suda ; Leslie A. Smith ; Livius V. d’Uscio ; Timothy E. Peterson ; and Zvonimir S. Katusic *

From the Departments of Anesthesiology, Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic College of Medicine, Rochester, Minn.

* To whom correspondence should be addressed. E-mail: katusic.zvonimir{at}mayo.edu.

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 (109) 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 p47phox 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.
Key words: endothelial function • endothelium • nitric oxide • nitric oxide synthase • reactive oxygen species




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