Thrombogenicity of the Injured Vessel Wall – Role of Antithrombin and Heparin

Summary

The thrombogenicity of the vessel wall after endothelial denudation is partly explained by an impaired inhibition of thrombin on the subendothelium. We have previously reported that thrombin coagulant activity can be detected on the vessel wall after balloon injury in vivo. The glycosaminoglycans of the subendothelium differ from those of the endothelium and have a lower catalyzing effect on antithrombin III, but inhibition of thrombin can still be augmented by addition of antithrombin III to the injured vessel surface.

In this study the effect of antithrombin III and heparin on thrombin coagulant activity on the vessel wall was studied after in vivo balloon injury of the rabbit aorta using biochemical and immunohistochemical methods and thrombin was analysed after excision of the vessel. Continuous treatment with heparin, lasting until sacrifice of the animal, or treatment with antithrombin III resulted in significant reduction of thrombin coagulant activity on the injured aorta. Heparin given only in conjunction with the injury did not prevent thrombin coagulant activity or deposition of fibrin on the surface.

The capacity of the injured vessel wall to inhibit thrombin in vitro was improved on aortic segments obtained from animals receiving antithrombin III but not from those given heparin. It is concluded that treatment with antithrombin III interferes with thrombin appearance on the vessel wall after injury and thereby reduces the risk for thrombosis.

• References

• 1 Owen WG, Esmon CT. Functional properties of an endothelial cell cofactor for thrombin-catalyzed activation of protein C. J Biol Chem 1981; 256: 5532-5
• 2 Esmon CT, Esmon NL, Harris KW. Complex formation between thrombin and thrombomodulin inhibits both thrombin-catalyzed fibrin formation and factor V activation. J Biol Chem 1982; 257: 7944-7
• 3 Marcum JA, Rosenberg RD. Anticoagulantly active heparin-like molecules from vascular tissue. Biochemistry 1984; 23: 1730-7
• 4 Rosenberg RD. Biochemistry of heparin antithrombin interactions, and the physiologic role of this natural anticoagulant mechanism. Am J Med 1989; 87: 2S-9S
• 5 De Bault LE, Esmon NL, Olson JR, Esmon CT. Distribution of the thrombomodulin antigen in the rabbit vasculature. Lab Invest 1986; 54: 172-8
• 6 Boffa M-C, Burke B, Haudenschild CC. Preservation of thrombomodulin antigen on vascular and extravascular surfaces. J Histochem Cytochem 1987; 35: 1267-76
• 7 Massaro TA, Glatz CE. Distribution of glycosaminoglycans in consecutive layers of the rabbit aorta. Artery 1979; 5: 1-13
• 8 Frebelius S, Nydahl S, Swedenborg J. Coagulant and noncoagulant thrombin enzymatic activity on the endothelium. Blood Coag Fibrinol 1990; 1: 285-92
• 9 Frebelius S, Swedenborg J. Uptake and inactivation of thrombin on the subendothelium: Comparisons with endothelium. Thromb Res 1987; 47: 223-33
• 10 Nydahl S, Frebelius S, Swedenborg J. Inhibition of thrombin on subendothelium. Studies on rabbit aorta, ex vivo. Eur Surg Res 1989; 21: 287-95
• 11 Groves HM, Kinlough-Rathbone RL, Richardson M, Moore S, Mustard JF. Platelet interaction with damaged rabbit aorta. Lab Invest 1979; 40: 194-200
• 12 Diyjski M, Olsson P, Swedenborg J. Thrombin activity appearing on the vessel wall after trauma. Thromb Haemostas 1985; 54: 773-5
• 13 Hatton MW C, Moar SL, Richardson M. Deendothelialization in vivo initiates a thrombogenic reaction at the rabbit aorta surface. Correlation of uptake of fibrinogen and antithrombin III with thrombin generation by exposed subendothelium. Am J Path 1989; 135: 499-508
• 14 Pasche B, Swedenborg J, Hedin U, Olsson P, Ljungqvist A. Thrombin-inhibitory capacity of the injured vessel wall. Thromb Res 1991; 62: 531-44
• 15 Chemnitz J, Christensen BC. Repair in arterial tissue. Demonstration of fibrinogen/fibrin in the normal and healing rabbit thoracic aorta by the indirect immunoperoxidase technique. Virchows Arch (A) 1984; 403: 163-71
• 16 Bylock A, Bondjers G. Early reactions of the arterial wall, following mechanical trauma. A scanning and transmission electron microscopy study. Acta Pathol Microbiol Scand (A) 1981; 89: 313-23
• 17 Nossel HL, Yudelman I, Canfield RE, Butler Jr VP, Spanondis K, Wilner GD, Qureshi GD. Measurement of fibrinopeptide A in human blood. J Clin Invest 1974; 54: 43-53
• 18 Kockum C, Frebelius S. Rapid radioimmunoassay of human fibrinopeptide A – removal of cross-reacting fibrinogen with bentonite. Thromb Res 1980; 19: 589-98
• 19 Swedenborg J, Dryjski M, Frebelius S, Olsson P. Uptake and inactivation of thrombin on rabbit aortic endothelium studied with two different substrates. Thromb Haemostas 1985; 54: 828-32
• 20 Abildgaard IJ, Lie M, Ødegaard OR. Antithrombin (heparin mfactor) assay with “new” chromogenic substrates (S-2238 and chromozym TH). Thromb Res 1977; 11: 549-53
• 21 Teien AN, Lie M, Abildgaard U. Assay of heparin in plasma using a chromogenic substrate for activated factor X. Thromb Res 1976; 8: 413-6
• 22 Kirk RE. Experimental Design: Procedures for the behavioral sciences. Brooks/Cole Publishing Company; Monterey: 1982: pp911
• 23 Busch C, Owen WG. Identification in vitro of an endothelial cell surface cofactor for antithrombin III. Parallel studies with isolated perfused rat hearts and microcarrier cultures of bovine endothelium. J Clin Invest 1982; 69: 726-9
• 24 Marcum JA, Mc Kenney JB, Rosenberg RD. Acceleration of thrombin-antithrombin complex formation in rat hindquarters via heparinlike molecules bound to the endothelium. J Clin Invest 1984; 74: 341-50
• 25 Shimada K, Ozawa T. The anticoagulant role of heparin-like molecules in the endothelial cell surface in hemostasis-endothelial heparinlike molecules. Acta Haematol Jpn 1986; 49: 1604-9
• 26 Pasche B, Swedenborg J, Frebelius S, Olsson P. Heparin cofactor II significance for the inhibition of thrombin at the injured vessel wall. Thromb Res 1991; 62: 409-19
• 27 de Agostini AI, Watkins SC, Slayter HS, Youssoufian H, Rosenberg RD. Localization of anticoagulantly active heparan sulfate proteoglycans in vascular endothelium: Antithrombin binding on cultured endothelial cells and perfused rat aorta. J Cell Biol 1990; 111: 1293-304
• 28 Wight TN. Cell biology of arterial proteoglycans. Arteriosclerosis 1989; 9: 1-20
• 29 Stemerman MB, Baumgartcr HR, Spaet TH. The subendothelial microfibrils and platelet adhesion. Lab Invest 1971; 24: 179-86
• 30 Groves HM, Kinlough-Rathbone RL, Richardson M, Jorgensen L, Moore S, Mustard JF. Thrombin generation and fibrin formation following injury to rabbit neointima. Studies of vessel wall reactivity and platelet survival. Lab Invest 1982; 46: 605-12
• 31 Zwaginga JJ, de Boer HC, Ijsseldijk MJ W, Kerkhof A, Miiller-Berghaus G, Gruhlichhenn JH, Sixma JJ, de Groot PG. Thrombogenicity of vascular cells. Comparison between endothelial cells isolated from different sources and smooth muscle cells and fibroblasts. Arteriosclerosis 1990; 10: 437-48
• 32 Baar-Shavit R, Eldor A, Vlodavsky I. Binding of thrombin to subendothelial extracellular matrix. Protection and expression of functional properties. J Clin Invest 1989; 84: 1096-104
• 33 Hatton MW C, Moar SL. Comparisons of the effects of heparin and hirudin on thrombin binding to the normal and the de-endothelialized rabbit aorta in vitro. Thromb Haemostas 1991; 66: 208-12
• 34 Hogg PJ, Jackson CM. Formation of a ternary complex between thrombin, fibrin monomer and heparin influences the action of thrombin on its substrates. J Biol Chem 1990; 265: 248-55
• 35 Hatton MW C, Moar SL, Richardson M. Evidence that rabbit ¹²⁵I-antithrombin III binds to proteoheparan sulfate at the subendothelium of the rabbit aorta in vitro. Blood Vessels 1988; 25: 12-27
• 36 Bini A, Fenoglio Jr J, Sobel J, Owen J, Fcjgl M, Kaplan KL. Immunochemical characterization of fibrinogen, fibrin I. and fibrin II in human thrombi and atherosclerotic lesions. Blood 1987; 69: 1038-45
• 37 Vu TH, Hung DT, Wheaton VI, Coughlin SR. The molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell 1991; 64: 1057-68
• 38 Sarembock IJ, Gertz SD, Gimple LW, Owen RM, Powers ER, Roberts WC. Effectiveness of recombinant desulpatohirudin in reducing restenosis after balloon angioplasty of atherosclerotic femoral arteries in rabbits. Circulation 1991; 84: 232-43
• 39 Hedin U, Frebelius S, Sanchez J, Dryjski M, Swedenborg J. Antithrombin III inhibits thrombin-induced proliferation in human arterial smooth muscle cells. Thromb Artcrioscl 1993. Accepted for publication