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Am J Physiol Heart Circ Physiol 283: H2560-H2566, 2002. First published August 22, 2002; doi:10.1152/ajpheart.00415.2002
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Vol. 283, Issue 6, H2560-H2566, December 2002

Inhibitory effect of recombinant iNOS gene expression on vasomotor function of canine basilar artery

Daihiko Eguchi1, Livius V. d'Uscio1, Chris Wambi1, Deborah Weiler1, Imre Kovesdi2, Timothy O'Brien3, and Zvonimir S. Katusic1

1 Departments of Anesthesiology and Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, Minnesota 55905; 2 Gene Vec, Incorporated, Gaithersburg, Maryland 20878; and 3 Department of Medicine, Clinical Sciences Institute, University College Hospital, Galway, Ireland

The present study was designed to determine the effect of recombinant inducible nitric oxide (NO) synthase (iNOS) gene expression on vasomotor function in cerebral arteries. Isolated canine basilar arteries were exposed ex vivo (30 min at 37°C) to an adenoviral vector [107, 108, or 109 plaque-forming units (pfu)/ml] encoding either the iNOS gene or the beta -galactosidase reporter gene. Twenty-four hours after transduction, Western blot analysis demonstrated expression of iNOS protein only in iNOS (109 pfu/ml)-transduced arteries. Immunohistochemical analysis localized iNOS expression predominantly in adventitia. Vascular reactivity of isolated basilar arteries was studied by isometric force recording. Concentration-response curves to UTP (10-9-10-3 M) and diethylaminodiazen-1-ium-1,2-dioate (10-10-10-5 M) were significantly shifted to the right in iNOS gene (109 pfu/ml)-transduced rings compared with control and beta -galactosidase-transduced rings (P < 0.05, n = 5-6). Endothelium-dependent relaxation to bradykinin was significantly attenuated in iNOS-transduced rings (P < 0.001, n = 8). The basal level of cGMP and superoxide anion (O<UP><SUB>2</SUB><SUP>−</SUP></UP>·) production were elevated in iNOS-transduced rings (P < 0.05, n = 7 for cGMP; P < 0.01, n = 6-9 for O<UP><SUB>2</SUB><SUP>−</SUP></UP>· production). Our results suggest that expression of recombinant iNOS in cerebral arteries reduces vasomotor reactivity to both vasoconstrictor and vasodilator agonists. Attenuation of contractions is most likely due to functional antagonism between UTP and cGMP. Reduction of endothelium-dependent relaxation to bradykinin appears to be mediated in part by reduced reactivity of smooth muscle cells to NO.

adventitia; bradykinin; nitric oxide; superoxide anion; inducible nitric oxide synthase


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