Effects of a Low Molecular Weight Heparin (Fragmin®) and of Unfractionated Heparin on Coagulation Activation at the Site of Plug Formation In Vivo

Sabine Elchinger; Michael Wolzt; Malgorzata Nieszpaur-Los; Barbara Schneider; Klaus Lechner; Hans-Georg Eichler; Paul A Kyrle

doi:10.1055/s-0038-1648970

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1994; 72(06): 831-835
DOI: 10.1055/s-0038-1648970

Original Article

Schattauer GmbH Stuttgart

Effects of a Low Molecular Weight Heparin (Fragmin®) and of Unfractionated Heparin on Coagulation Activation at the Site of Plug Formation In Vivo

Sabine Elchinger

The Department of Medicine I, Division of Hematology and Hemostaseology, Department of Clinical Pharmacology, and Institute of Medical Statistics and Documentation, Vienna University Hospital, Vienna, Austria

,

Michael Wolzt

The Department of Medicine I, Division of Hematology and Hemostaseology, Department of Clinical Pharmacology, and Institute of Medical Statistics and Documentation, Vienna University Hospital, Vienna, Austria

,

Malgorzata Nieszpaur-Los

The Department of Medicine I, Division of Hematology and Hemostaseology, Department of Clinical Pharmacology, and Institute of Medical Statistics and Documentation, Vienna University Hospital, Vienna, Austria

,

Barbara Schneider

The Department of Medicine I, Division of Hematology and Hemostaseology, Department of Clinical Pharmacology, and Institute of Medical Statistics and Documentation, Vienna University Hospital, Vienna, Austria

,

Klaus Lechner

The Department of Medicine I, Division of Hematology and Hemostaseology, Department of Clinical Pharmacology, and Institute of Medical Statistics and Documentation, Vienna University Hospital, Vienna, Austria

,

Hans-Georg Eichler

The Department of Medicine I, Division of Hematology and Hemostaseology, Department of Clinical Pharmacology, and Institute of Medical Statistics and Documentation, Vienna University Hospital, Vienna, Austria

,

Paul A Kyrle

The Department of Medicine I, Division of Hematology and Hemostaseology, Department of Clinical Pharmacology, and Institute of Medical Statistics and Documentation, Vienna University Hospital, Vienna, Austria

› Author Affiliations

Abstract

PDF Download

Summary

The clinical benefits of unfractionated heparin (UFH) and low molecular weight heparin (LMWH) have been shown in many trials. However, the mode of action of heparin has not been fully elucidated. Thus, we wanted to study the effects of UFH and LMWH in vivo by measuring coagulation activation markers in blood obtained directly from a vascular injury site. In a double-blind, randomized, 3-way, cross-over study 18 healthy volunteers were given UFH (150 U/kg s.c.) and 2 doses of LMWH [35 U/kg s.c. (low dose, Id), 75 U/kg s.c. (high dose, hd)]. Prothrombin fragment 1+2 (F₁₊₂), thrombin-antithrombin complex (TAT) and fibrinopeptide A (FPA) were measured in bleeding time blood and in venous blood before and after drug application. In addition, the effects of UFH and LMWH on in vitro coagulation tests were studied. Compared to base line, UFH and both IdLMWH and hdLMWH caused significant reductions of F₁₊₂, TAT and FPA in bleeding time blood at 2 h. A marked effect of UFH and of hdLMWH was also seen at 5 h. The inhibition of FPA generation was more pronounced after hdLMWH compared to IdLMWH. In venous blood, UFH and LMWH caused reductions of F₁₊₂, but not of TAT and FPA. In vitro, UFH predominantly affected the anti-IIa assays (activated partial thromboplastin time, thrombin time) and LMWH mainly the anti-Xa test system. Using a technique that investigates the activated coagulation system in vivo, a time- and dose dependent inhibitory effect of heparin on coagulation activation was detectable. Therefore, in our experimental setting a preferential inhibition of a particular portion of the coagulation system by one of the two heparin preparations was not detectable.

PDF (386 kb)

References

References
1 Barrowcliffe TW, Johnson EA, Thomas DP. Low Molecular Weight Heparin. Chichester, UK: Wiley and Sons; 1992
2 Thomgren M, Shafi S, Born GVR. Thromboxane A₂ in skin-bleeding time blood and in clotted venous blood before and after administration of acetyl-salicylic acid. Lancet 1983; 1: 1075-1078
3 Kyrle PA, Westwick J, Scully MF, Kakkar VV, Lewis GP. Investigation of the interaction of blood platelets with the coagulation system at the site of plug formation in vivo in man - effect of low-dose aspirin. Thromb Haemost 1987; 57: 62-66
4 Weiss HJ, Lages B. Evidence for tissue factor-dependent activation of the classic extrinsic coagulation mechanism in blood obtained from bleeding time wounds. Blood 1988; 71: 629-635
5 Gerrard JM, Taback S, Singhroy S, Docherty JC, Kostolansky I, McNicol A, Kobrinsky NL, McKenzie JK, Rowe R. In vivo measurement of thromboxane B₂ and 6-keto-prostaglandin F_1α in humans in response to a standardized vascular injury and the influence of aspirin. Circulation 1989; 79: 29-38
6 Nowak J, FitzGerald GA. Redirection of prostaglandin endoperoxide metabolism at the platelet-vascular interface in man. J Clin Invest 1989; 83: 380-305
7 Gresele P, Arnout J, Deckmyn H, Huybrechts E, Pieters G, Vermylen J. Role of proaggregatory and antiaggregatory prostaglandins in hemostasis -Studies with combined thromboxane synthase inhibition and thromboxane receptor antagonism. J Clin Invest 1987; 80: 1435-1445
8 Bauer KA. Laboratory markers of coagulation activation. Arch Pathol Lab Med 1993; 117: 71-77
9 Pelzer H, Schwart A, Stuber W. Determination of human thrombin-anti-thrombin complex in plasma with an enzyme-linked immunoabsorbent assay. Thromb Haemost 1988; 59: 101-106
10 Nossel H, Younger L, Wilner G, Procupez T, Canfiled R, Butler VpJr. Radioimmunoassay of human fibrinopeptide A. Proc Natl Acad Sci USA 1971; 68: 2350-2353
11 Mielke CH, Kaneshiro MM, Mahler IA, Weiner JM. Rapaport SI. The standardized normal Ivy bleeding time and its prolongation by aspirin. Blood 1969; 34: 204-215
12 Barrowcliffe TW, Le Shirley Y. The effect of calcium chloride on anti-Xa activity of heparin and its molecular weight fractions. Thromb Haemost 1989; 62: 950-954
13 Lane DA, Pejler G, Flynn AM, Thompson EA, Lindahl U. Neutralization of heparin-related saccharides by histidin-rich glycoprotein and platelet factor 4. J Biol Chem 1986; 261: 3980-3986
14 Michalski R, Lane DA, Pepper DS, Kakkar VV. Neutralisation of heparin in plasma by platelet factor four and protamine sulphate. Br J Haem 1978; 38: 561-571
15 Sandset M, Abildgaard U, Larsen ML. Heparin induces release of extrinsic coagulation activation pathway inhibitor (EPI). Thromb Res 1988; 50: 803-813
16 Lindahl AK, Abildgaard U, Larsen ML, Aamodt LM, Nordfang O, Beek TC. Extrinsic pathway inhibitor (EPI), and the post-heparin anticoagulant effect in tissue thromboplastin-induced coagulation. Thromb Res 1991; (Suppl. 14) 39-48
17 Dawes J, Pumphrey CW, McLaren KM, Prowse CV, Pepper DS. The in vivo release of human platelet factor 4 by heparin. Thromb Res 1982; 27: 65-76
18 O’Brien JR, Etherington MD, Pashley MA. The heparin mobilisable pool of platelet factor 4: a comparison of intravenous and subcutaneous heparin and Kabi heparin fragment 2165. Thromb Haemost 1985; 54: 735-738
19 Barrowcliffe TW, Eggleton CA, Stocks J. Studies of anti-Xa activity in human plasma. II: The role of lipoproteins. Thromb Res 1982; 27: 185-195
20 Barrowcliffe TW, Havercroft SJ, Kemball-Cook G, Lindahl U. The effect of Ca²⁺, phospholipids and factor V on the anti-(factor Xa) activity of heparin and its high-affinity oligosaccharides. Biochem J 1987; 243: 31-37
21 Ellis V, Scully MF, Kakkar VV. The acceleration of the inhibition of platelet prothrombinase complex by heparin. Biochem J 1986; 233: 161-165
22 Green D, Hirsh J, Heit J, Prins M, Davidson B, Lensing AWA. Low molecular weight: a critical analysis of clinical trials. Pharmacol Rev 1994; 461: 89-109
23 Johnson EA, Kirkwood TBL, Stirling Y, Perez-Requejo JL, Ingram GIC, Bangham DR, Brozovic M. Four heparin preparations: anti-Xa potentiating effect of heparin after subcutaneous injection. Thromb Haemost 1975; 35: 586-591
24 Brandt JT. Heparin anticoagulation. In: Practical laboratory hematology Koepke JA. ed. Churchill Livingstone, New York, Edinburgh, London, Melbourne, Tokyo: 1991: 433-454
25 Bauer K, Mannucci PM, Gringeri A, Tradati F, Barzegar S, Kass B, ten Cate H, Kestin A, Brettler D, Rosenberg R. Factor IXa-factor VIIIa-cell surface complex does not contribute to the basal activation of the coagulation mechanism in vivo. Blood 1992; 79: 2039-2047
26 Radcliffe R, Nemerson Y. Activation and control of factor VII by activated factor X and thrombin. J Biol Chem 1975; 250: 388-395
27 Gailani D, Broze G. Factor XI activation by thrombin and factor XIa. Sem Throm Hem 1993; 19: 396-404
28 Broze G. The role of tissue factor pathway inhibitor in a revised coagulation cascade. Sem Hem 1992; 29: 159-169
29 Ozge-Anwar AH, Conell GE, Mustard JF. The activation of factor VIII by thrombin. Blood 1965; 26: 500-509
30 Suzuki K, Dahlbäck B, Stenflo J. Thrombin-catalyzed activation of human coagulation factor V. J Biol Chem 1982; 257: 6556-6564