An NADPH oxidase superoxide-generating system in the rabbit aorta.
Superoxide anion can modulate vascular smooth muscle tone and potentially affect the growth response in vascular disease. The present studies were undertaken to characterize the source of superoxide in rabbit aorta. Rings of aorta (5 mm) were incubated in physiological salt solution (PSS) for 30 min at 37 degrees C in the presence of 10 mM diethyldithiocarbamate (DDC) with or without inhibitors of superoxide-generating systems. Rings were then placed in PSS containing 250 microM lucigenin at 37 degrees C in the presence or absence of inhibitors, and changes in amounts of superoxide were determined by measuring chemiluminescence (units). The inhibitors of xanthine oxidase, oxypurinol (300 microM), and of mitochondrial NADH dehydrogenase, rotenone (50 microM), had no significant effect on superoxide levels. An inhibitor of NADPH oxidase, iodonium thiophen, caused a concentration-dependent inhibition of superoxide anion (12.49 +/- 1.48 vs 5.27 +/- 1.81 and 2.30 +/- 0.36 units, control vs 7 microM and 70 microM iodonium thiopen, respectively). A structurally related iodonium compound, diphenyleneiodonium (20 microM), caused a 78% reduction in basal and DDC-evoked superoxide levels. In the presence or absence of DDC, exogenous administration of NADPH (10 microM-1 mM), but not NADP (1 mM), elicited a concentration-dependent rise in superoxide levels that was inhibited by iodonium thiophen. Particulate fractions of whole aortic tissue exhibited NADPH-dependent superoxide production that was inhibited by 1 microM diphenyleneiodonium.(ABSTRACT TRUNCATED AT 250 WORDS)
Pagano
P J
PJ
Robert Dawson Evans Department of Clinical Research, University Medical Center, Boston 02118, USA.
Ito
Y
Y
Tornheim
K
K
Gallop
P M
PM
Tauber
A I
AI
Cohen
R A
RA
D000818:Animals; D001013:Aorta, Thoracic / Q000201:enzymology*; D004050:Ditiocarb / Q000494:pharmacology; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000201:enzymology*; D009247:NADH, NADPH Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D019255:NADPH Oxidases; D009861:Onium Compounds / Q000494:pharmacology; D010117:Oxypurinol / Q000494:pharmacology; D011817:Rabbits; D012402:Rotenone / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology; D013481:Superoxides / Q000378:metabolism*; D013876:Thiophenes / Q000494:pharmacology
D009861:Onium Compounds; D013876:Thiophenes; D012402:Rotenone; D013481:Superoxides; C051503:iodonium thiophene; D009249:NADP; C007517:diphenyleneiodonium; D004050:Ditiocarb; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; D010117:Oxypurinol