title: Antithrombotic action of annexin V proved as efficient as direct inhibition of tissue factor or thrombin.
abstract: The role of phospholipid platelet membrane and tissue factor in thrombin generation and thrombus formation is accepted. In the present study we have explored antithrombotic action of strategies aimed to block exposure of negatively charged phospholipids and we compared effects with those obtained through tissue factor or a direct thrombin inhibition. Type III collagen was exposed to flowing blood (5 min, 300 s(-1)). Effects of inhibition of platelet deposition by annexin A5 (ANXA5), hirudin (HIR) or by an antibody against tissue factor (TF) were evaluated. Prothrombin fragment F1 + 2 (F1 + 2) was monitored. Pre-incubation of whole blood with HIR or ANXA5 resulted in a statistically significant reduction of platelet deposition (12.2 +/- 0.6% in control experiments vs. 8.3 +/- 0.4% and 8.5 +/- 0.5%, respectively, P < 0.05). A similar decrease was found when blood was incubated with an antibody against TF. Furthermore, ANXA5 and HIR inhibited the recruitment of platelets into forming aggregates. The height of platelet aggregates generated was decreased in the presence of HIR or ANXA5, but only incubation with both inhibitors reached levels of statistical significance. The presence of ANXA5 or HIR decreased levels of F1 + 2 suggesting a reduced activation of the coagulation system. In our experimental studies, the inhibitory potential of ANXA5 on platelet-thrombus formation was as effective as that of a direct thrombin inhibitor, as HIR, or an antibody against TF. Negatively charged phospholipids exposed on activated platelets potentiate the formation of platelet aggregates on a collagen surface and further suggest that inhibition of platelet procoagulant activity might be a specific target for antithrombotic drugs.
authors.affiliation: Servicio de Hemoterapia y Hemostasia. Hospital Clínic, CDB, IDIBAPS, Facultad de Medicina, Barcelona, Spain. agalan@clinic.ub.es
authors.forename: A M
authors.initials: AM
authors.lastname: Galán
authors.forename: W L
authors.initials: WL
authors.lastname: van Heerde
authors.forename: G
authors.initials: G
authors.lastname: Escolar
authors.forename: A
authors.initials: A
authors.lastname: Ordinas
authors.forename: J
authors.initials: J
authors.lastname: Sixma
authors.forename: P G
authors.initials: PG
authors.lastname: de Groot
mesh_terms: D017304:Annexin A5 / Q000378:metabolism / Q000494:pharmacology*; D000906:Antibodies / Q000276:immunology; D001792:Blood Platelets / Q000187:drug effects* / Q000276:immunology; D002462:Cell Membrane / Q000378:metabolism; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D006629:Hirudins / Q000494:pharmacology; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods; D010446:Peptide Fragments / Q000032:analysis; D010974:Platelet Aggregation / Q000187:drug effects / Q000276:immunology; D011498:Protein Precursors / Q000032:analysis; D011516:Prothrombin / Q000032:analysis; D013925:Thromboplastin / Q000276:immunology
chemical_list: D017304:Annexin A5; D000906:Antibodies; D005343:Fibrinolytic Agents; D006629:Hirudins; D010446:Peptide Fragments; D011498:Protein Precursors; C029733:prothrombin fragment 1; C044183:prothrombin fragment 2; D011516:Prothrombin; D013925:Thromboplastin