Inhibition of clot lysis and decreased binding of tissue-type plasminogen activator as a consequence of clot retraction.
Tissue-type plasminogen activator (t-PA) is less active in vivo and in vitro against clots that are enriched in platelets, even at therapeutic concentrations. The release of radioactivity from 125I-fibrin-labeled clots was decreased by 47% 6 hours after the addition of t-PA 400 U/mL when formed in platelet-rich versus platelet-poor plasma. This difference was not due to the release of plasminogen activator inhibitor-1 (PAI-1) by platelets. Thus, the fibrinolytic activity of t-PA in the supernatant was similar in the two preparations and fibrin autography demonstrated only a minor degree of t-PA-PAI-1 complex formation. Furthermore, a similar platelet-dependent reduction in clot lysis was seen with a t-PA mutant resistant to inhibition by PAI-1. The reduction in t-PA activity correlated with a decrease in t-PA binding to platelet-enriched clot (60% +/- 3% v platelet-poor clot, n = 5). This reduction in binding was also shown using t-PA treated with the chloromethylketone, D-Phe-Pro-Arg-CH2Cl (PPACK) (36% +/- 13%, n = 3), and with S478A, a mutant t-PA in which the active site serine at position 478 has been substituted by alanine (43% +/- 6%, n = 3). In contrast, fixed platelets and platelet supernatants had no effect on the binding or lytic activity of t-PA. Pretreatment with cytochalasin D 1 mumol/L, which inhibits clot retraction, also abolished the platelet-induced inhibition of lysis and t-PA binding by platelets. These data suggest that platelets inhibit clot lysis at therapeutic concentrations of t-PA as a consequence of clot retraction and decreased access of fibrinolytic proteins.
Kunitada
S
S
Division of Clinical Pharmacology, Vanderbilt University, Nashville, TN.
FitzGerald
G A
GA
Fitzgerald
D J
DJ
D001792:Blood Platelets / Q000502:physiology; D003018:Clot Retraction; D005337:Fibrin / Q000378:metabolism; D005342:Fibrinolysis; D006801:Humans; D010958:Plasminogen / Q000378:metabolism; D015849:Plasminogen Inactivators / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000378:metabolism*
D015849:Plasminogen Inactivators; D010980:Platelet Membrane Glycoproteins; D005337:Fibrin; D010958:Plasminogen; D010959:Tissue Plasminogen Activator