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Thromb Haemost 1994; 71(05): 655-662
DOI: 10.1055/s-0038-1642499
Review Article
Schattauer GmbH Stuttgart

Clopidogrel – a Platelet Inhibitor which Inhibits Thrombogenesis in Non-Anticoagulated Human Blood Independently of the Blood Flow Conditions

Helge E Roald
1   The Biotechnology Centre of Oslo, University of Oslo, Oslo, Norway
,
R Marius Barstad
1   The Biotechnology Centre of Oslo, University of Oslo, Oslo, Norway
,
Peter Kierulf
2   The Clinical Chemistry Department, Ullevaal University Hospital, Oslo, Norway
,
Fredrik Skjørten
3   The Department of Pathology, Ullevaal University Hospital, Oslo, Norway
,
John P Dickinson
4   The Sanofi Winthrop Ltd., Manchester, UK
,
Gilles Kieffer
5   The Sanofi Recherche, Montpellier, France
,
Kjell S Sakariassen
1   The Biotechnology Centre of Oslo, University of Oslo, Oslo, Norway
› Author Affiliations
Further Information

Publication History

Received 05 October 1993

Accepted after revision 19 January 1993

Publication Date:
06 July 2018 (online)

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Summary

The goal of the present study was to investigate the effect of 7 and 14 days of daily oral administration of 75 mg clopidogrel on collagen-induced thrombogenesis in flowing non-anticoagulated human blood. Blood was drawn directly from an antecubital vein over immobilised collagen type III fibrils positioned in a parallel-plate perfusion chamber. The wall shear rates at the collagen surface were those characteristic for veins (100 s−1), and for medium sized (650 s−1) and moderately stenosed (2600 s−1) arteries.

Clopidogrel ingestion reduced the thrombus volume significantly (p <0.05) at 100 and 2600 s−1 (39 and 51% respectively). The β-thromboglobulin plasma levels were reduced concomitantly. However, it was not possible to measure accurately the thrombus volume at 650 s−1, due to loose packing of the platelet thrombi. Transmission electron micros-copy substantiated this observation and showed that clopidogrel profoundly reduced the platelet degranulation process (p <0.005). The inhibitory effect of clopidogrel on platelet consumption by the growing thrombi resulted apparently in higher platelet concentration at the collagen surface, which enhanced the platelet-collagen adhesion at all three shear rates (p <0.05). Despite the low deposition of fibrin on collagen, clopidogrel reduced significantly the fibrinopeptide A plasma levels and the fibrin deposition at shear rates below 650 s−1. This was apparently a consequence of the reduced platelet recruitment and the lower activation of platelets, since activated platelets in thrombi promote deposition of fibrin.

Thus, it appears that clopidogrel inhibits thrombus formation in human blood primarily by interfering with platelet-platelet binding and the inhibition is effective independently of the wall shear rate.

 

  • References

  • 1 Packham MA, Mustard JF. The role of platelets in the development and complications of atherosclerosis. Semin Hematol 1986; 23: 8-26
    PubMedGoogle Scholar
  • 2 Hennekens CH, Peto R, Hutchinson GB, Doll R. An overview of the British and American aspirin studies. N Engl J Med 1988; 318: 923-4
    CrossrefPubMedGoogle Scholar
  • 3 Yusuf S, Wittes L, Friedmann L. Overview of results of randomized clinical trials in heart disease. II. Unstable angina, heart failure, primary prevention with aspirin and risk factor modification. JAMA 1988; 260: 2259-63
    CrossrefPubMedGoogle Scholar
  • 4 Lewis HD, Davis JW, Archibald DG, Steinke WE, Smitherman TC, Doherty JE, Schnaper HW, Le Winter MM, Linares E, Pouget JM, Sabharwal SC, Chesler E, De Mots H. Protective effects of aspirin against acute myocardial infarction and death in men with unstable angina. N Engl J Med 1983; 309: 396-403
    CrossrefPubMedGoogle Scholar
  • 5 Carins JA, Gent M, Singer J, Finnie KJ, Froggatt GM, Holder DA, Jablonsky G, Kostuk P, Melendez LJ, Myers MG, Sackett DL, Sealey BJ, Tanser PH. Aspirin sulfinpyrazone or both in unstable angina: Results of a Canadian multicenter trial. N Engl J Med 1985; 313: 1369-75
    CrossrefPubMedGoogle Scholar
  • 6 Théroux P, Quimet H, Mc Cans J, Latour JG, Joly P, Lévy G, Pelletier E, Juneau M, Stasiak J, de Guise P, Pelletier GB, Rinzler D, Waters DD. Aspirin, heparin, or both to treat unstable angina. N Engl J Med 1989; 319: 1105-11
    PubMedGoogle Scholar
  • 7 The RISC Group. Risk of myocardial infarction and death during treatment with low dose aspirin and intravenous heparin in men with unstable coronary artery disease. Lancet 1990; 336: 827-30
    CrossrefPubMedGoogle Scholar
  • 8 Mills DCB, Puri R, Hu CJ, Minniti C, Grana G, Freedman MD, Freedman S, Colman RF, Colman RW. Clopidogrel inhibits the binding of ADP analogues to the receptor mediating inhibition of platelet adenylate cyclase. Arterioscler Thromb 1992; 12: 430-6
    CrossrefPubMedGoogle Scholar
  • 9 Féliste R, Delebassée D, Simon MF, Chap H, Defreyn G, Vallée E, Douste-Blazy L, Maffrand JP. Broad spectrum antiplatelet activity of Ticlopidine and PCR 4099 involves the suppression of the effects of released ADP. Thromb Res 1987; 48: 403-15
    CrossrefPubMedGoogle Scholar
  • 10 Féliste R, Simon MF, Chap H, Douste-Blazy L, Defreyn G, Maffrand JP. Effect of PCR 4099 on ADP-induced calcium movements and phosphatidic acid production in rat platelets. Biochem Pharm 1988; 37: 2559-64
    CrossrefPubMedGoogle Scholar
  • 11 Sakariassen KS, Joss R, Muggli R, Kuhn H, Tschopp T, Sage H, Baumgartner HR. Collagen type III induced ex vivo thrombogenesis in humans: role of platelets and leukocytes in deposition of fibrin. Arteriosclerosis 1990; 10: 276-84
    CrossrefPubMedGoogle Scholar
  • 12 Roux SP, Sakariassen KS, Turitto VT, Baumgartner HR. Effect of aspirin and epinephrine on experimentally induced thrombogenesis in dogs. A parallelism between in vivo and ex vivo thrombosis models. Arteriosclerosis Thrombosis 1991; 11: 1182-91
    CrossrefPubMedGoogle Scholar
  • 13 Fressinaud E, Sakariassen KS, Turitto VT, Baumgartner HR. Shear rate-dependent impairment of thrombus growth on collagen in nonanticoagulated blood from patients with von Willebrand disease and haemophilia A. Blood 1992; 80: 988-94
    CrossrefPubMedGoogle Scholar
  • 14 Sakariassen KS, Cousot D, Hadváry P, Baumgartner HR. Aspirin ingestion reduces thrombus volume in human nonanticoagulated blood only at shear rates characteristic for stenosed arteries. Thromb Haemost 1991; 65: 782 (Abstr)
    PubMedGoogle Scholar
  • 15 Roald HE, Barstad RM, Engen A, Kierulf P, Skjprten F, Sakariassen KS. HN-11500 – a novel thromboxane A, receptor antagonist with antithrombotic activity in humans at arterial blood flow conditions. Thromb Haemost 1994; 71: 103-9
    Article in Thieme ConnectPubMedGoogle Scholar
  • 16 Baumgartner HR. Effects of anticoagulation on the interaction of human platelets with subendothelium in flowing blood. Schweiz Med Wochenschr 1976; 106: 1367-8
    PubMedGoogle Scholar
  • 17 Sakariassen KS, Aarts PAMM, de Groot PG, Houdijk WPM, Sixma JJ. A perfusion chamber developed to investigate platelet interaction in flowing blood with human vessel wall cells, their extracellular matrix and purified components. J Lab Clin Med 1983; 102: 522-35
    PubMedGoogle Scholar
  • 18 Miller EF, Rhodes RK. Preparation and characterization of the different types of collagen. Methods Enzymol 1982; 82: 3-64
    PubMedGoogle Scholar
  • 19 Burns RR, Gross J. High resolution analysis of the modified quarter-stagger model of the collagen fibril. Biopolymers 1977; 13: 931-41
    PubMedGoogle Scholar
  • 20 Sakariassen KS, Muggli R, Baumgartner HR. Measurements of platelet interaction with components of the vessel wall in flowing blood. Methods Enzymol 1989; 169: 37-70
    PubMedGoogle Scholar
  • 21 Sakariassen KS, Weiss HJ, Baumgartner HR. Upstream thrombus growth impairs downstream thrombogenesis in non-anticoagulated blood: effect of procoagulant artery subendothelium and non-procoagulant collagen. Thromb Haemost 1991; 65: 596-600
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 Baumgartner HR, Muggli R. Adhesion and aggregation: morphological demonstration and quantitation in vivo and in vitro. In: Platelets in Biology and Pathology. Gordon JL. (ed) Elsevier, Amsterdam; The Netherlands: 1976: 23-60
    Google Scholar
  • 23 Sakariassen KS, Kuhn H, Muggli R, Baumgartner HR. Growth and stability of thrombi in flowing citrated blood: Assessment of platelet surface interactions with computer-assisted morphometry. Thromb Haemost 1988; 60: 392-8
    Article in Thieme ConnectPubMedGoogle Scholar
  • 24 Ludlam CA, Cash JL. Studies on the liberation of β-llirornboglobulin from human platelets in vitro. Brit J Haematol 1976; 3: 239-47
    PubMedGoogle Scholar
  • 25 Sakariassen KS, Nicvclstcin PFEM, Collcr B, Sixma JJ. The role of platelet membrane glycoprotein Ib and IIb/IIIa in platelet adherence to human artery subendothelium. Brit J Haematol 1986; 63: 681-91
    CrossrefPubMedGoogle Scholar
  • 26 Nurden AT, Caen JP. An abnormal platelet glycoprotein pattern in three cases of Glanzmann’s thrombastenia. Br J Haematol 1974; 28: 253-60
    CrossrefPubMedGoogle Scholar
  • 27 Sakariassen KS, Roald HE, Salatti JA. Ex vivo models for studying thrombosis: special emphasis on shear rate dependent blood-collagen interactions. In: Advances in Cardiovascular Engineering. Hwang NHC, Turitto VT, Yen MRT. (eds) New York: Plenum Press; 1992: 151-74
    Google Scholar
  • 28 Miletich JP, Jackson CM, Majerus PW. Properties of the factor Xa binding site on human platelets. J Biol Chem 1978; 253: 6908-16
    PubMedGoogle Scholar
  • 29 Rosing J, Bevers EM, Confurius P, Hemker HC, van Diejen G, Weiss HJ, Zwaal RFA. Impaired factor X and prothrombin activation associated with decreased phospholipid exposure in platelets from a patient with a bleeding disorder. Blood 1985; 65: 1557-61
    PubMedGoogle Scholar
  • 30 Tracy PB. Regulation of thrombin generation at cell surfaces. Sem Thromb Haemostas 1988; 14: 227-33
    PubMedGoogle Scholar
  • 31 Weiss HJ, Turitto VT, Baumgartner HR. Role of shear rate and platelets in promoting fibrin formation on rabbit subendothelium. Studies utilizing patients with quantitative and qualitative platelet defects. J Clin Invest 1986; 78: 1072-82
    CrossrefPubMedGoogle Scholar
  • 32 Wilcox JN, Smith KM, Schwartz SM, Gordon D. Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci USA 1989; 86: 2889-93
    PubMedGoogle Scholar
  • 33 Drake TA, Morrissey JH, Edginglon TS. Selective cellular expression of tissue factor in human tissue. Implications for disorders of hemostasis and thrombosis. Am J Pathol 1989; 134: 1087-97
    PubMedGoogle Scholar