Inhibition of concomitant thrombin-mediated fibrin formation enhances clot lysis in whole blood.
To characterize potential mechanisms for enhanced thrombolysis in the presence of thrombin inhibitors, we measured lysis of 125I-fibrin-(ogen)- labelled clots after incubation in rotating tubes with whole blood containing tissue-type plasminogen activator (t-PA, 1500 ng/ml) and either saline or increasing concentrations of recombinant desulphatohirudin (hirudin), or heparin. Thrombin activity in washed clots was negligible, but incubation of clots with t-PA in non-anticoagulated blood resulted in marked thrombin activity within 5 min as measured by fibrinopeptide A generation. Incubation with hirudin over a range of concentrations increased clot lysis compared with t-PA alone (control) from 132 +/- 18% of control with 0.5 microgram/ml (n = 4) to 216 +/- 48% of control with 10 micrograms/ml (n = 4). Incubation with less than 0.5 U/ml of heparin attenuated clot lysis (35 +/- 22% of control with 0.08 U/ml, n = 3) while concentrations > or = 0.5 U/ml increased lysis (178 +/- 13% of control with 1.7 U/ml, n = 4). Similar results were obtained for incubations in recalcified platelet-poor plasma indicating that platelets are not required for the enhancement of clot lysis induced by thrombin inhibitors. However, incubations in recalcified plasma from a patient with afibrinogenaemia abolished the increased lysis observed with 10 micrograms/ml of hirudin and addition of physiological concentrations of fibrin monomer (400-800 nM) or fibrinogen degradation products (500 nM) to the mixture of whole blood, t-PA and hirudin blunted the extent of clot lysis. Thus, inhibition of thrombin activity induced by exposure of clots to t-PA prevents concomitant formation of fibrin that attenuates fibrinolysis.(ABSTRACT TRUNCATED AT 250 WORDS)
Haskel
E J
EJ
Cardiovascular Division, Washington University School of Medicine, St Louis, MO 63110.
Eisenberg
P R
PR
Abendschein
D R
DR
D001769:Blood / Q000187:drug effects*; D005342:Fibrinolysis / Q000187:drug effects*; D006493:Heparin / Q000494:pharmacology* / Q000627:therapeutic use; D006629:Hirudins / Q000494:pharmacology*; D006801:Humans; D007457:Iodine Radioisotopes; D011994:Recombinant Proteins / Q000494:pharmacology; D013917:Thrombin / Q000037:antagonists & inhibitors*; D015912:Thrombolytic Therapy; D010959:Tissue Plasminogen Activator / Q000494:pharmacology
D006629:Hirudins; D007457:Iodine Radioisotopes; D011994:Recombinant Proteins; D006493:Heparin; D013917:Thrombin; D010959:Tissue Plasminogen Activator