title: Inhibition of the prothrombinase complex on red blood cells by heparin and covalent antithrombin-heparin complex.
abstract: The role of red blood cells (RBCs) in coagulation is not well understood. Overt exposure of phosphatidylserine on surfaces of RBCs provide docking sites for formation of the prothrombinase complex, which further aids in amplification of coagulation leading to subsequent thrombosis. No studies to date have evaluated heparin inhibition of the RBC-prothrombinase system. Therefore, this study examines the ability of heparin and a covalent antithrombin-heparin complex (ATH) to inhibit the RBC-prothrombinase system. Discontinuous inhibition assays were performed to obtain k₂ values for inhibition of free or prothrombinase-bound Xa by antithrombin and unfractionated heparin (AT + UFH) versus ATH. In addition, components of the complex (prothrombin, RBCs or Va) were excluded prior to reaction with inhibitors to investigate potential mechanisms involved. Inhibition of thrombin generation, fibrinogen conversion and plasma clotting by the RBC-prothrombinase system was also examined. Protection of Xa was observed for AT + UFH and not for ATH reactions. Inhibition rates for ATH were significantly faster when compared with AT + UFH results. The greatest impact on Xa inhibition was observed from factor Va omission for both inhibitors. ATH inhibited thrombin generation, fibrinogen conversion and plasma clotting better compared with AT + UFH. This study determined potential control of coagulation contributed by RBCs. Moreover, greater control of coagulation is achieved by covalently linking heparin to AT.
authors.affiliation: Thrombosis and Atherosclerosis Research Institute (TaARI), DBCVSRI, Hamilton General Hospital, 237 Barton Street East, Hamilton, ON, L8L 2X2, Canada.
authors.forename: Ivan
authors.initials: I
authors.lastname: Stevic
authors.forename: Howard H W
authors.initials: HH
authors.lastname: Chan
authors.forename: Leslie R
authors.initials: LR
authors.lastname: Berry
authors.forename: Ankush
authors.initials: A
authors.lastname: Chander
authors.forename: Anthony K C
authors.initials: AK
authors.lastname: Chan
mesh_terms: D000328:Adult; D000925:Anticoagulants / Q000737:chemistry / Q000494:pharmacology*; D000991:Antithrombins / Q000737:chemistry / Q000494:pharmacology*; D001777:Blood Coagulation / Q000187:drug effects; D004912:Erythrocytes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D005165:Factor V / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015951:Factor Xa / Q000378:metabolism; D065427:Factor Xa Inhibitors; D005337:Fibrin / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D006493:Heparin / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013450:Sulfones / Q000494:pharmacology
chemical_list: D000925:Anticoagulants; D000991:Antithrombins; D065427:Factor Xa Inhibitors; D015842:Serine Proteinase Inhibitors; D013450:Sulfones; C022475:prothrombinase complex; C002010:4-(2-aminoethyl)benzenesulfonylfluoride; D005165:Factor V; D005337:Fibrin; D005340:Fibrinogen; D006493:Heparin; D015951:Factor Xa