Enhanced binding of fibrinogen by the subendothelium after treatment of the rabbit aorta with thrombin.
Damage to the endothelial surface of an artery invokes a hemostatic response that causes platelet deposition and activation of the coagulation and fibrinolytic pathways on the exposed subendothelial surface. Plasma fibrinogen is rapidly adsorbed at the site of injury. To gain insight into fibrinogen uptake, undamaged and de-endothelialized rabbit thoracic aortas were pretreated with various concentrations of thrombin and then incubated with fibrinogen labeled with iodine 125 in vitro. Uptake of fibrinogen by the subendothelium was not affected by low thrombin concentrations (less than 10 nmol/L), probably because of the antithrombin capacity of the vessel wall to inactivate any thrombin adsorbed. Over the thrombin concentration range of 10 to 90 nmol/L. fibrinogen binding increased linearly as binding of thrombin labeled with iodine 131 increased. In contrast, treatment of the subendothelium with enzymatically inactive thrombin did not enhance fibrinogen binding. Fibrinogen binding was inhibited by exposing the thrombin-treated subendothelium to hirudin or phenylalanyl-prolyl-arginyl-chloromethyl ketone. High thrombin concentrations (greater than 100 nmol/L) caused either a steadily decreasing uptake of fibrinogen with low fibrinogen concentrations or fibrin coagulation on the subendothelial surface from a high fibrinogen concentration. Glycyl-prolyl-arginyl-proline (0.1 mg/ml), a selective inhibitor of fibrin polymerization, inhibited 72% to 78% of fibrinogen uptake by the thrombin-treated subendothelium. Fibrinogen uptake was Ca2(+)-dependent, but ethylenediaminetetraacetic acid (10 mmol/L) did not displace subendothelium-bound fibrinogen. Plasmin effectively removed at least 75% of bound fibrinogen, indicating an extracellular location for the protein.(ABSTRACT TRUNCATED AT 250 WORDS)
Hatton
M W
MW
Department of Pathology, McMaster University Health Sciences Centre, Hamilton, Ontario, Canada.
Moar
S L
SL
Richardson
M
M
D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000378:metabolism*; D001485:Basement Membrane / Q000378:metabolism; D001665:Binding Sites / Q000187:drug effects; D001692:Biological Transport / Q000187:drug effects; D002118:Calcium / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D005340:Fibrinogen / Q000378:metabolism*; D005341:Fibrinolysin / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D066298:In Vitro Techniques; D009842:Oligopeptides / Q000494:pharmacology; D011817:Rabbits; D013917:Thrombin / Q000494:pharmacology*
D009842:Oligopeptides; C034585:glycyl-prolyl-arginyl-proline; D005340:Fibrinogen; D006493:Heparin; C046617:arginyl-glycyl-aspartyl-serine; D013917:Thrombin; D005341:Fibrinolysin; D002118:Calcium