Matrix metalloproteinase-2 in platelet adhesion to fibrinogen: interactions with nitric oxide.
BACKGROUND
Matrix metalloproteinase-2 (MMP-2) has been shown to activate a non-thromboxane, non-ADP pathway of platelet aggregation. In contrast, nitric oxide (NO) is known to inhibit platelet adhesion and aggregation. Therefore, we have studied the release of MMP-2 during platelet adhesion to fibrinogen, the effects of phenanthrolione, an MMP-2 inhibitor, on adhesion and the interactions of inhibitor with a NO donor, S-nitroso-N-acetyl-D, L-penicillamine (SNAP).


METHODS
Human platelets were isolated from blood of healthy volunteers and platelet adhesion to fibrinogen-coated plates was studied by measuring thrombin-stimulated release of platelet a-granule constituent, platelet factor 4. In addition, the mode of action of phenanthroline and NO on platelets was investigated by assaying the levels of intraplatelet cyclic GMP.


RESULTS
Thrombin-stimulated platelet adhesion to fibrinogen was associated with increased release of MMP-2 from platelets. Phenanthroline (0.1-100 KM) reduced platelet adhesion to fibrinogen. The adhesion was also inhibited by SNAP (0.1-100 KM), an effect abolished by 1H-[1,2,4] oxadiazolol [4,3,-a] quinoxalin-1-one (ODQ), a selective inhibitor of the soluble guanylate cyclase. Co-administration of phenanthroline and SNAP resulted in a synergistic inhibition of platelet adhesion, an effect that was not associated with enhanced cyclic GMP generation by platelets. Furthermore, ODQ did not reverse the synergistic effect of these compounds on adhesion.


CONCLUSIONS
1. MMP-2 promotes platelet adhesion to fibrinogen. 2. Phenanthroline and NO synergize to inhibit platelet adhesion to fibrinogen acting through a cyclic GMP-independent mechanism(s).
Martinez
A
A
Department of Pharmacology, University of Valencia, Valencia, Spain.
Salas
E
E
Radomski
A
A
Radomski
M W
MW
D001792:Blood Platelets / Q000201:enzymology / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D006152:Cyclic GMP / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D010069:Oxadiazoles / Q000494:pharmacology; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D010618:Phenanthrolines / Q000494:pharmacology; D010974:Platelet Aggregation; D011810:Quinoxalines / Q000494:pharmacology; D013917:Thrombin / Q000378:metabolism
C095284:1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one; D004791:Enzyme Inhibitors; D020030:Nitric Oxide Donors; D010069:Oxadiazoles; D010618:Phenanthrolines; D011810:Quinoxalines; C110881:S-nitro-N-acetylpenicillamine; D009569:Nitric Oxide; D005340:Fibrinogen; D013917:Thrombin; D020778:Matrix Metalloproteinase 2; D010396:Penicillamine; D006152:Cyclic GMP; C025205:1,10-phenanthroline