Improvement of relaxation in an atherosclerotic artery by gene transfer of endothelial nitric oxide synthase.
Gene transfer with replication-deficient adenovirus is a useful tool to study vascular biology. We have reported that overexpression of endothelial nitric oxide (NO) in carotid arteries from normal rabbits augments vasorelaxation mediated by NO. In this study, we tested the hypothesis that adenovirus-mediated gene transfer of endothelial nitric oxide synthase (eNOS) improves impaired relaxation of atherosclerotic vessels. We used 2 replication-deficient adenoviruses: AdeNOS, which carries cDNA for eNOS, and Adbetagal, which expresses beta-galactosidase. Common carotid arteries from 10 New Zealand White (NZW; plasma cholesterol, 79+/-13 mg/dL) and 10 Watanabe heritable hyperlipidemic (WHHL; plasma cholesterol, 452+/-39 mg/dL) rabbits were incubated in organ culture with AdeNOS, Adbetagal, or vehicle alone. Carotid arteries from WHHL rabbits had mild to moderate atherosclerotic lesions. Histochemical staining for beta-galactosidase and immunohistochemistry for eNOS indicated transgene expression in the endothelium and adventitia in both NZW and WHHL rabbits. Expression of eNOS determined with Western blot analysis after incubation with AdeNOS tended to be higher in vessels from WHHL rabbits than NZW rabbits. Effects of transgene expression on vascular function were examined by recording isometric tension 1 day after transduction. After precontraction with phenylephrine, acetylcholine produced significantly less relaxation in vessels from WHHL rabbits than in vessels from NZW rabbits. Relaxation in response to acetylcholine was greater in carotid arteries from both NZW and WHHL rabbits that were transfected with AdeNOS than in vessels treated with vehicle or Adbetagal. Vasorelaxation in response to acetylcholine was inhibited by Nomega-nitro-L-arginine. Responses to sodium nitroprusside were similar after treatment with vehicle alone, Adbetagal, or AdeNOS in both groups of rabbits. Thus, overexpression of eNOS with an adenoviral vector improves impaired NO-mediated relaxation in atherosclerotic arteries.
Ooboshi
H
H
Departments of Internal Medicine and Pharmacology (F.M.F., D.D.H.), Cardiovascular Center and Center on Aging, University of Iowa College of Medicine, Iowa City, USA.
Toyoda
K
K
Faraci
F M
FM
Lang
M G
MG
Heistad
D D
DD
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001161:Arteriosclerosis / Q000628:therapy*; D015153:Blotting, Western; D017536:Carotid Artery, Common / Q000503:physiopathology; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D018014:Gene Transfer Techniques; D006651:Histocytochemistry; D008297:Male; D019001:Nitric Oxide Synthase / Q000235:genetics*; D052250:Nitric Oxide Synthase Type III; D011817:Rabbits; D015398:Signal Transduction / Q000502:physiology; D014664:Vasodilation / Q000502:physiology*; D014779:Virus Replication / Q000235:genetics; D001616:beta-Galactosidase / Q000032:analysis
D018076:DNA, Complementary; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; D001616:beta-Galactosidase