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Thromb Haemost 1987; 58(02): 744-748
DOI: 10.1055/s-0038-1645978
Original Articles
Schattauer GmbH Stuttgart

Effect of Dipyridamole on Spontaneous Platelet Aggregation in Whole Blood Decreases with the Time After Venepuncture: Evidence for the Role of ADP

A R Saniabadi
The Haemostasis and Thrombosis Research Unit, University Department of Medicine, Royal Infirmary, Glasgow, UK
,
G D O Lowe
The Haemostasis and Thrombosis Research Unit, University Department of Medicine, Royal Infirmary, Glasgow, UK
,
J C Barbenel
*   The Bioengineering Unit, University of Strathclyde, Glasgow, UK
,
C D Forbes
The Haemostasis and Thrombosis Research Unit, University Department of Medicine, Royal Infirmary, Glasgow, UK
› Author Affiliations
Further Information

Publication History

Received 30 December 1986

Accepted after revision 02 April 1987

Publication Date:
27 June 2018 (online)

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Summary

Spontaneous platelet aggregation (SPA) was studied in human whole blood at 3, 5, 10, 20, 30, 40 and 60 minutes after venepuncture. Using a whole blood platelet counter, SPA was quantified by measuring the fall in single platelet count upon rollermixing aliquots of citrated blood at 37° C. The extent of SPA increased with the time after venepuncture, with a correlation coefficient of 0.819. The inhibitory effect of dipyridamole (Dipy) on SPA was studied: (a) 10 μM at each time interval; (b) 0.5-100 μM at 3 and 30 minutes and (c) 15 μM in combination with 100 μM adenosine, 8 μM 2-chloroadenosine (2ClAd, an ADP receptor blocker) and 50 μM aspirin. There was a rapid decrease in the inhibitory effect of Dipy with the time after venepuncture; the correlation coefficient was -0.533. At all the concentrations studied, Dipy was more effective at 3 minutes than at 30 minutes after venepuncture. A combination of Dipy with adenosine, 2ClAd or aspirin was a more effective inhibitor of SPA than either drug alone. However, when 15 μM Dipy and 10 μM Ad were added together, the inhibitory effect of Dipy was not increased significantly, suggesting that Dipy inhibits platelet aggregation independent of Ad. The increase in SPA with the time after venepuncture was abolished when blood was taken directly into the anticoagulant containing 5 μM 2ClAd. It is suggested that ADP released from the red blood cells is responsible for the increased platelet aggregability with the time after venepuncture and makes a serious contribution to the artifacts of in vitro platelet function studies.

Keywords

Time after venepuncture - Spontaneous platelet aggregation - Dipyridamole - 2-chloroadenosine - ADP
 

  • References

  • 1 Weiss HJ. Dipyridamole and other phosphodiesterase inhibitors. In Platelets Pathophysiology and Antiplatelet Drug Therapy. Harvey JWeiss. (Ed), pp 57-62 Alan R Liss Inc; New York: 1982
    Google Scholar
  • 2 Harker LA, Kadatz RA. Mechanisms of action of dipyridamole. Thromb Res 1983; Suppl IV 39-46
    PubMedGoogle Scholar
  • 3 Gresele P, Zoja C, Deckmyn H, Amout J, Vermylen J, Verstraete M. Dipyridamole inhibits platelet aggregation in whole blood. Thromb Haemostas 1983; 50: 852-856
    PubMedGoogle Scholar
  • 4 Kincaird-Smith P. Modification of vascular lesions of rejections in cadaveric renal allografts by dipyridamole and anticoagulants. Lancet 1969; 2: 920-922
    PubMedGoogle Scholar
  • 5 Harker LA, Slinchter SJ. Studies of platelet and fibrinogen kinetics in patients with prosthetic heart valves. N Engl J Med 1970; 283: 1302-1305
    CrossrefPubMedGoogle Scholar
  • 6 Sullivan JM, Harken DW, Gorlin R. Pharmacologic control of thromboembolic complications of cardiac valve replacement. N Engl J Med 1971; 284: 1391-1394
    CrossrefPubMedGoogle Scholar
  • 7 Persantin-aspirin Reinfarction Study Research Group Persantin and aspirin in coronary heart disease. Circulation 1980; 62: 449-461
    CrossrefPubMedGoogle Scholar
  • 8 Chesebro JH, Clements IP, Fuster V, Elveback LR, Smith HC, Bardsley WT, Frye RL, Holmes DR, Vliestra RE, Pluth JR, Wallace RB, Puga FJ, Orszulak TA, Piehler JM, Schaff HV, Danielson GK. A platelet inhibitory drug trial in coronary artery bypass operations: benefit of preoperative dipyridamole and aspirin therapy on early postoperative vein graft patency. N engl J Med 1982; 307: 73-78
    CrossrefPubMedGoogle Scholar
  • 9 Chesebro JH, Fuster V, Elveback LD, Elveback LR, Clements IP, Smith HC, Holmes DR, Bardsley WT, Pluth JR, Wallace RB, Puga JF, Orszulak TA, Piehler JM, Danielson GK, Schaff HV, Frye RL. Effect of dipyridamole and aspirin on late vein graft patency after coronary bypass operations. N Engl J Med 1984; 310: 209-214
    CrossrefPubMedGoogle Scholar
  • 10 Best LC, McGuire MB, Jones P BB, Holand TK, Marti J, Preston FE, Segat DS, Russell R GG. Mode of action of dipyridamole on human platelets. Thromb Res 1979; 16: 367-379
    CrossrefPubMedGoogle Scholar
  • 11 Smith JB, Mills D CB. Inhibition of adenosine 3’-5’-cyclic monophosphate diesterase. Biochem J 1970; 120: 20p
    PubMedGoogle Scholar
  • 12 Rozenberg MC, Walker CM. The effect of pyrimidine compounds on the potentiation of adenosine inhibition of aggregation, on adenosine phosphorylation and phosphodiesterase activity of blood platelets. Br J Haematol 1973; 24: 409-418
    CrossrefPubMedGoogle Scholar
  • 13 Emmons PR, Harrison M GJ, Honour AJ, Mitchell J RA. Effect of pyrimidopyrimidine derivative on thrombus formation in the rabbit. Nature 1965; 208: 255-257
    CrossrefPubMedGoogle Scholar
  • 14 Harker LA, Hanson SR. Experimental arterial thromboembolism in baboons: mechanisms, quantitation and pharmacological prevention. J Clin Invest 1979; 64: 559-569
    CrossrefPubMedGoogle Scholar
  • 15 Born G VR. Quantitative investigation into the aggregation of blood platelets. J Physiol 1962; 67-68
    PubMedGoogle Scholar
  • 16 Harrison M GJ, Pollock SS, Steiner M, Weisblatt E. Inhibitors of spontaneous platelet aggregation in whole blood. Atherosclerosis 1985; 58: 199-203
    CrossrefPubMedGoogle Scholar
  • 17 Heptinstal S, Fox S, Crawford J, Hawkins M. Inhibition of platelet aggregation in whole blood by dipyridamole and aspirin. Thromb Res 1986; 42: 215-223
    CrossrefPubMedGoogle Scholar
  • 18 Santos MT, Aznar J, Valles J, Perez-Requejo JL. Evaluation in whole blood of antiplatelet activity of dipyridamole and pentoxifylline with the basic wave. Thromb Res 1986; Suppl IV: Abstr 296
    PubMedGoogle Scholar
  • 19 Saniabadi AR, Lowe G DO, Barbenel JC, Forbes CD. A comparison of spontaneous platelet aggregation in whole blood with platelet rich plasma: additional evidence for the role of ADP. Thromb Haemostas 1984; 51: 115-118
    PubMedGoogle Scholar
  • 20 Saniabadi AR, Lowe G DO, Barbenel JC, Forbes CD. Further studies on the role of red blood cells in spontaneous platelet aggregation. Thromb Res 1985; 38: 225-232
    CrossrefPubMedGoogle Scholar
  • 21 Haslam RJ, Lynham JA. Activation and inhibition of blood platelet adenylate cyclase by adenosine and 2-chloroadenosine. Life Sci 1972; 11: 1143-1154
    CrossrefPubMedGoogle Scholar
  • 22 Premont J, Preez M, Bockaert J. Adenosine-sensitive adenylate cyclase in rate striatal homogenates (Letter). FEBS 1977; 75: 209-212
    CrossrefPubMedGoogle Scholar
  • 23 Nachman RL, Ferris B. Binding of adenosine diphosphate by isolated membranes from human platelets. J Biol Chem 1974; 249: 704-710
    PubMedGoogle Scholar
  • 24 Gaarder A, Jonsen J, Laland S, Hellem A, Owren PA. Adenosine diphosphate in red cells as a factor in the adhesiveness of human blood platelets. Nature 1961; 192: 531-532
    CrossrefPubMedGoogle Scholar
  • 25 Born G VR. Arterial thrombosis and its prevention. In: proceedings of the VIII World Congress of Cardiology. Shoji H, Satoru M. (Eds) pp 81-91 Excerpta Medica; Amsterdam. Oxford: 1979
    Google Scholar
  • 26 Aursnes I, Gjesdai K, Abildgaard U. Platelet aggregation by ADP from unsheared erythrocytes at physiological Ca++ concentration. Br J Haematol 1981; 47: 149-152
    CrossrefPubMedGoogle Scholar
  • 27 Fox SC, Bergess-Wilson M, Heptinstal S, Mitchell J RA. Platelet aggregation in whole blood determined using the Ultra Flo 100 platelet counter. Thromb Haemostas 1983; 48: 327-329
    PubMedGoogle Scholar
  • 28 Saniabadi AR, Lowe G DO, Belch J JF, Forbes CD, Prentice C RM, Barbenel JC. The novel effects of a new prostacyclin analogue ZK36374 on the aggregation of human platelets in whole blood. Thromb Haemostas 1983; 50: 718-721
    PubMedGoogle Scholar
  • 29 Saniabadi AR, Lowe G DO, Belch J JF, Barbenel JC, Forbes CD. Effect of prostacyclin (Epoprostenol) on the aggregation of human platelets in whole blood in vitro. Haemostasis 1984; 14: 487-494
    PubMedGoogle Scholar
  • 30 Moncada S, Korbut R. Dipyridamole and other phosphodiesterase inhibitors act as antithrombotic agents by potentiating endogenous prostacyclin. Lancet 1978; 1: 1286-1289
    PubMedGoogle Scholar
  • 31 Steer ML, MacIntyre DE, Levine L, Salzman EW. Is prostacyclin a physiologically important circulating antiplatelet agent. Nature 1980; 283: 194-195
    CrossrefPubMedGoogle Scholar
  • 32 Terres W, Becker BF, Kratzer M AA, Gerlach E. Increased platelet aggregability after venepuncture is platelet not plasma derived. Thromb Res 1986; 539-548
    PubMedGoogle Scholar
  • 33 Gryglewski RJ. Prostacyclin and atherosclerosis. Trend Pharmacol Sci 1980; 2: 164-166
    PubMedGoogle Scholar