NAD(P)H oxidase-dependent platelet superoxide anion release increases platelet recruitment.
Platelets, although not phagocytotic, have been suggested to release O. Since O-producing reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) oxidases can be specifically activated by certain agonists and are found in several nonphagocytotic tissues, we investigated whether such an enzyme is the source of platelet-derived O. We further studied which agonists cause platelet O release and whether platelet-derived O influences thrombus formation in vitro. Collagen, but not adenosine 5'-diphosphate (ADP) or thrombin, increased O formation in washed human platelets. This was a reduced nicotinamide adenine dinucleotide (NADH)-dependent process, as shown in platelet lysates. Consistent with a role of a platelet, NAD(P)H oxidase expression of its subunits p47(phox) and p67(phox) and inhibition of platelet O formation by diphenylene-iodoniumchloride (DPI) and by the specific peptide-antagonist gp91ds-tat were observed. Whereas platelet-derived O did not influence initial aggregation, platelet recruitment to a preformed thrombus following collagen stimulation was significantly attenuated by superoxide dismutase (SOD) or DPI. It was also inhibited when ADP released during aggregation was cleaved by the ectonucleotidase apyrase. ADP in supernatants of collagen-activated platelets was decreased in the presence of SOD, resulting in lower ADP concentrations available for recruitment of further platelets. Exogenous O increased ADP- concentrations in supernatants of collagen-stimulated platelets and induced irreversible aggregation when platelets were stimulated with otherwise subthreshold concentrations of ADP. These results strongly suggest that collagen activation induces NAD(P)H oxidase-dependent O release in platelets, which in turn enhances availability of released ADP, resulting in increased platelet recruitment.
Krötz
Florian
F
Institute of Physiology and the Department of Internal Medicine, Ludwig-Maximilians-University Munich, Schillerstr 44, 80336 Munich, Germany. fkroetz@lmu.de
Sohn
Hae Young
HY
Gloe
Torsten
T
Zahler
Stefan
S
Riexinger
Tobias
T
Schiele
Thomas M
TM
Becker
Bernhard F
BF
Theisen
Karl
K
Klauss
Volker
V
Pohl
Ulrich
U
D000244:Adenosine Diphosphate / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D003094:Collagen / Q000494:pharmacology; D004347:Drug Interactions; D006801:Humans; D007700:Kinetics; D008954:Models, Biological; D019255:NADPH Oxidases / Q000378:metabolism / Q000502:physiology*; D010974:Platelet Aggregation / Q000187:drug effects*; D013482:Superoxide Dismutase / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism* / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology; D013927:Thrombosis / Q000209:etiology / Q000473:pathology
D013481:Superoxides; D000244:Adenosine Diphosphate; D003094:Collagen; D013482:Superoxide Dismutase; D019255:NADPH Oxidases; D013917:Thrombin