Nitric oxide released by platelets inhibits neutrophil B2 integrin function following acute carbon monoxide poisoning.
Carbon monoxide (CO) poisoning has been reported to temporarily inhibit B2 integrin adherence molecules on leukocytes in previous studies in a rat model. The aim of this study was to investigate the mechanism for this effect. Studies were conducted using blood obtained from rats after they were exposed to CO and also with blood cells exposed to CO in vitro. Initial investigations indicated that inhibition of neutrophil (polymorphonuclear leukocyte, PMN) B2 integrin function was linked to the platelets in blood, as the effect was resolved by decreasing platelet number before PMN adherence was tested. The platelet effect could also be shown by incubating either platelet-rich plasma or whole blood with CO in vitro. The effect of platelets was blocked by superoxide radicals and by NG-nitro-L-arginine methyl ester, an inhibitor of nitric oxide (NO) synthase. These observations suggested that CO caused platelets to release NO, an agent known to inhibit the function of B2 integrins. The concentration of NO measured in suspensions of platelets from rats poisoned with CO according to the established model (exposure to 1000 ppm CO for 40 min and 3000 ppm CO for 20 min) was 47 nmol/10(8) platelets, in contrast to only 0.3 nmol NO/10(8) platelets from control rats. Enhanced NO release occurred despite a 60% inhibition of NO synthase activity, assessed by measuring conversion of [14C]L-arginine to citrulline. Exposure to only 1000 ppm CO for 1 hr caused platelets to release 74 nmol NO/10(8) platelets, and no inhibition of NO synthase occurred. Enhanced NO release, and inhibition of PMN adherence, did not occur after platelets were exposed to light from a quartz lamp to photodissociate CO from heme proteins. The data suggest that the NO flux from platelets increased when CO became bound to heme-containing platelet proteins, which normally scavage intraplatelet NO and thus prevent diffusion beyond the platelet membrane.
Thom
S R
SR
Institute for Environmental Medicine, University of Pennsylvania, Philadelphia 19104.
Ohnishi
S T
ST
Ischiropoulos
H
H
D000594:Amino Acid Oxidoreductases / Q000378:metabolism; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D002249:Carbon Monoxide Poisoning / Q000097:blood / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D004347:Drug Interactions; D016023:Integrins / Q000378:metabolism*; D008297:Male; D019331:NG-Nitroarginine Methyl Ester; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D009569:Nitric Oxide / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000494:pharmacology; D019001:Nitric Oxide Synthase; D051381:Rats; D017208:Rats, Wistar
D016023:Integrins; D009569:Nitric Oxide; D001120:Arginine; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; D019331:NG-Nitroarginine Methyl Ester