title: Resistance of endothelium-dependent relaxation to elevation of O(-)(2) levels in rabbit carotid artery.
abstract: 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 microM; 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.
authors.affiliation: Vascular Biology Unit, Boston Medical Center, Boston, Massachusetts 02118, USA. ppagano1@hfhs.org
authors.forename: P J
authors.initials: PJ
authors.lastname: Pagano
authors.forename: M C
authors.initials: MC
authors.lastname: Griswold
authors.forename: S
authors.initials: S
authors.lastname: Najibi
authors.forename: S L
authors.initials: SL
authors.lastname: Marklund
authors.forename: R A
authors.initials: RA
authors.lastname: Cohen
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D001012:Aorta, Abdominal / Q000187:drug effects / Q000378:metabolism; D001013:Aorta, Thoracic / Q000187:drug effects / Q000378:metabolism; D002339:Carotid Arteries / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D004050:Ditiocarb / Q000494:pharmacology; D004351:Drug Resistance; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D011817:Rabbits; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D014664:Vasodilation / Q000187:drug effects / Q000502:physiology*
chemical_list: D004791:Enzyme Inhibitors; D007527:Isoenzymes; D009569:Nitric Oxide; D004050:Ditiocarb; D013482:Superoxide Dismutase; D000109:Acetylcholine; D010100:Oxygen; D019331:NG-Nitroarginine Methyl Ester