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Am J Physiol Heart Circ Physiol 277: H2109-H2114, 1999;
0363-6135/99 $5.00
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Vol. 277, Issue 5, H2109-H2114, November 1999

Resistance of endothelium-dependent relaxation to elevation of Ominus 2 levels in rabbit carotid artery

Patrick J. Pagano1, Mark C. Griswold1, Soheil Najibi1, Stefan L. Marklund2, and Richard A. Cohen1

1 Vascular Biology Unit, Boston Medical Center, Boston, Massachusetts 02118; and 2 Department of Clinical Chemistry, Umeå University Hospital, Umeå, Sweden

Endogenous superoxide anion (O-2) interferes with the bioactivity of nitric oxide (NO) in endothelium-dependent arterial relaxation (EDR). Using the lucigenin chemiluminescence assay, we measured O-2 in the thoracic and abdominal aortas and the carotid artery of rabbits to determine whether ambient O-2 varies among the three arteries and differentially diminishes the effect of NO. Basal levels of O-2 were significantly higher in carotid arteries than in the thoracic aorta [23 ± 6.1 vs. 3.9 ± 1.4 chemiluminescence units (CU); P < 0.05], whereas EDR in response to ACh (10-8-10-5 M) was not significantly different on ANOVA. After treatment with the superoxide dismutase (SOD) inhibitor diethyldithiocarbamate (DDC; 10 mM), O-2 levels were significantly elevated, becoming greater in the carotid artery and abdominal aorta than in the thoracic aorta (185 ± 31.2 and 202 ± 40.3 vs. 89 ± 18 CU; P < 0.05). DDC significantly reversed EDR in the thoracic aorta but not in the carotid artery; at 10-6 M ACh, the decrease seen with DDC was 48 ± 6.2 vs. 6.8 ± 8.0% of maximal relaxation in the thoracic aorta and carotid artery, respectively. In the thoracic aorta, exogenous SOD reversed the inhibition of EDR caused by DDC. Moreover, DDC/O-2-resistant EDR in the carotid artery was ablated by the addition of nitro-L-arginine methyl ester (300 µM; P < 0.05), an NO synthase inhibitor, consistent with peroxynitrite or an O-2-resistant NO donor being involved in carotid relaxation. Indeed, exogenous peroxynitrite caused similar relaxation of the carotid artery and thoracic aorta, which was unaffected by DDC. Our studies show a greater production of nitrite and O-2 per unit area by the carotid artery, suggesting a greater amount of their product peroxynitrite. These findings support the hypothesis that peroxynitrite is the relaxing agent that resists high O-2 in the carotid artery.

aorta; reactive oxygen species; superoxide anion


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