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Thromb Haemost 1998; 80(04): 662-667
DOI: 10.1055/s-0037-1615439
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Schattauer GmbH

Ethanol Inhibits Thrombin-induced Secretion of the Contents of Human Platelet Dense and α-Granules and Lysosomes

A. Nguyen
1   From the Department of Biochemistry, Toronto, Ontario, Canada
,
C. H. Gemmell
2   From the Department of Medicine, University of Toronto, Toronto, Ontario, Canada
,
E. L. Yeo
2   From the Department of Medicine, University of Toronto, Toronto, Ontario, Canada
,
M. A. Packham
1   From the Department of Biochemistry, Toronto, Ontario, Canada
,
M. L. Rand
1   From the Department of Biochemistry, Toronto, Ontario, Canada
3   From the Division of Haematology/Oncology, The Hospital for Sick Children, Toronto, Ontario, Canada
› Author Affiliations
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Publication History

Received 26 February 1998

Accepted after resubmission 17 June 1998

Publication Date:
08 December 2017 (online)

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Summary

Moderate consumption of alcoholic beverages is associated with a reduction in thromboembolic complications of coronary artery disease, possibly partially attributable to inhibition by ethanol of platelet responses to some aggregating agents. Although ethanol is known to inhibit thrombin-induced secretion of platelet dense granule contents, the effect of ethanol on secretion of α-granule and lysosomal contents has not been studied. Using suspensions of washed platelets, and a range of thrombin concentrations (up to 0.1 U/ml), we examined the effect of 87 mM ethanol on secretion of [14C]serotonin from prelabelled platelets as a measure of secretion of dense granule contents. Secretion of α-granule and lysosomal contents was examined by flow cyto-metric measurement of the surface expression of CD62P (P-selectin) and CD63, respectively. Secretion of the lysosomal enzyme, β-N-acetylglucosaminidase was also quantified. Results were expressed as % of maximum response induced by 1 U/ml thrombin. Ethanol inhibited the thrombin-induced secretion of both dense and α-granule contents (P <0.001, 2-way ANOVA), and of lysosomal contents (P <0.005 for CD63 expression and P <0.001 for β-N-acetylglucosaminidase secretion). When platelets were pretreated with aspirin, thrombin-induced secretion of storage granule and lysosomal contents was slightly inhibited, but secretion was inhibited by ethanol to the same extent as the untreated platelets, indicating that this inhibition was independent of thromboxane A2. Surface expression of CD63 occurred at lower thrombin concentrations than those required for secretion of β-N-acetylglucosaminidase, possibly due to the presence of some CD63 on granule membranes. Although the role of lysosomal contents in thrombus formation is not established, some constituents of storage granules are known to augment thrombus formation; ethanol’s inhibition of their secretion by stimulated platelets may contribute to its beneficial effect on thromboembolism.

Presented in part at the XVIth Congress of the International Society on Thrombosis and Haemostasis, Florence, Italy, June 1997.

 

  • References

  • 1 Rimm EB, Giovannucci EL, Willett WC, Colditz GA, Ascherio A, Rosner B, Stampfer MJ. Prospective study of alcohol consumption and risk of coronary disease in men. Lancet 1991; 338: 464-8.
    CrossrefPubMedGoogle Scholar
  • 2 Klatsky AL, Armstrong MA, Friedman GD. Risk of cardiovascular mortality in alcohol drinkers, ex-drinkers and non-drinkers. Am J Cardiol 1990; 66: 1237-42.
    CrossrefPubMedGoogle Scholar
  • 3 Kannel WB, Ellison RC. Alcohol and coronary heart disease: The evidence for a protective effect. Clin Chim Acta 1996; 246: 59-76.
    CrossrefPubMedGoogle Scholar
  • 4 Moore RD, Pearson TA. Moderate alcohol consumption and artery disease. Medicine 1986; 65: 242-67.
    CrossrefPubMedGoogle Scholar
  • 5 Rubin R, Rand ML. Alcohol and platelet function. Alcoholism: Clin Exp Res 1994; 18: 105-10.
    CrossrefPubMedGoogle Scholar
  • 6 Latta EK, Packham MA, DaCosta SM, Rand ML. Effects of chronic administration of ethanol on platelets from rabbits with diet-induced hyper-cholesterolemia. Unchanged characteristics and responses to ADP but reduction of enhanced thrombin-induced, TxA2-independent platelet responses. Arterioscler Thromb 1994; 14: 1372-7.
    CrossrefPubMedGoogle Scholar
  • 7 Latta EK, Packham MA, Gross PL, Rand ML. Enhanced collagen-induced responses of platelets from rabbits with diet-induced hypercholesterolemia are due to increased sensitivity to TxA2. Response inhibition by chronic ethanol administration in hypercholesterolemia is due to reduced TxA2 formation. Arterioscler Thromb 1994; 14: 1379-85.
    CrossrefPubMedGoogle Scholar
  • 8 Rand ML, Groves HM, Packham MA, Mustard JF, Kinlough-Rathbone RL. Acute administration of ethanol to rabbits inhibits thrombus formation induced by indwelling aortic catheters. Lab Invest 1990; 63: 742-5.
    PubMedGoogle Scholar
  • 9 Mikhailidis DP, Jeremy JY, Barradas MA, Green N, Dandona P. Effect of ethanol on vascular prostacyclin (prostaglandin I2) synthesis, platelet aggregation, and platelet thromboxane release. Br Med J 1983; 287: 1495-8.
    CrossrefPubMedGoogle Scholar
  • 10 Rand ML, Groves HM, Kinlough-Rathbone RL, Packham MA, Mustard JF. Effects of ethanol on rabbit platelet responses to collagen in vitro and ex vivo and on platelet adhesion to de-endothelialized aortae in vivo. Thromb Res 1987; 48: 379-88.
    CrossrefPubMedGoogle Scholar
  • 11 Rand ML, Packham MA, Kinlough-Rathbone RL, Mustard JF. Effects of ethanol on pathways of platelet aggregation in vitro. Thromb Haemost 1988; 59: 383-7.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 12 Rubin R. Ethanol interferes with collagen-induced platelet activation by inhibition of arachidonic acid mobilization. Arch Biochem Biophys 1989; 270: 99-113.
    CrossrefPubMedGoogle Scholar
  • 13 Rand ML, Vickers JD, Kinlough-Rathbone RL, Packham MA, Mustard JF. Thrombin-induced inositol trisphosphate production by rabbit platelets is inhibited by ethanol. Biochem J 1988; 251: 279-84.
    CrossrefPubMedGoogle Scholar
  • 14 Packham MA. Role of platelets in thrombosis and hemostasis. Can J Physiol Pharmacol 1994; 72: 278-84.
    CrossrefPubMedGoogle Scholar
  • 15 Berman CL, Yeo EL, Wencel-Drake JD, Furie BC, Ginsberg MH, Furie B. A platelet alpha granule membrane protein that is associated with the plasma membrane after activation. J Clin Invest 1986; 78: 130-7.
    CrossrefPubMedGoogle Scholar
  • 16 Michelson AD. Flow cytometry: A clinical test of platelet function. Blood 1996; 87: 4925-36.
    PubMedGoogle Scholar
  • 17 van Oost BA. In Vitro Platelet Responses: Acid Hydrolase Secretion. In: Platelet Responses and Metabolism. Volume 1: Responses. Holmsen H, ed. Boca Raton, Florida: CRC Press; 1986: 163-91.
    Google Scholar
  • 18 Nieuwenhuis HK, van Oosterhout JJG, Rozemuller E, van Iwaarden F, Sixma JJ. Studies with a monoclonal antibody against activated platelets: evidence that a secreted 53,000-molecular weight lysosome-like granule protein is exposed on the surface of activated platelets in the circulation. Blood 1987; 70: 838-45.
    PubMedGoogle Scholar
  • 19 Chronos NAF, Wilson DJ, Janes SL, Hutton RA, Buller NP, Goodall AH. Aspirin does not affect the flow cytometric detection of fibrinogen binding to, or release of α-granules or lysosomes from, human platelets. Clin Sci 1994; 87: 575-80.
    CrossrefPubMedGoogle Scholar
  • 20 Taylor ML, Misso NLA, Stewart GA, Thompson PJ. Differential expression of platelet activation markers CD62P and CD63 following stimulation with PAF, arachidonic acid and collagen. Platelets 1995; 6: 394-401.
    CrossrefPubMedGoogle Scholar
  • 21 Koike K, Holmsen H. Requirement for thrombin receptor occupancy during platelet secretion under aggregating and nonaggregating conditions. Thromb Haemost 1987; 58: 1053-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 Molnar J, Lorand L. Studies on apyrases. Arch Biochem Biophys 1961; 93: 353-63.
    CrossrefPubMedGoogle Scholar
  • 23 Kinlough-Rathbone RL, Packham MA, Mustard JF. Platelet Aggregation. In: Methods in Hematology. Measurements of Platelet Function. Harker LA, Zimmerman TS, eds. Edinburgh: Churchill-Livingstone; 1983: 64-91.
    Google Scholar
  • 24 Aster RH, Jandl JH. Platelet sequestration in man. I. Methods. J Clin Invest 1964; 43: 843-55.
    CrossrefPubMedGoogle Scholar
  • 25 Rand ML, DaCosta SM, Chahil A, Kinlough-Rathbone RL. Combined inhibitory effects of ethanol and adenosine on the responses of rabbit platelets to thrombin. Alcoholism: Clin Exp Res 1995; 19: 1279-85.
    CrossrefPubMedGoogle Scholar
  • 26 Holmsen H, Dangelmaier CA. Measurement of secretion of lysosomal acid glycosidases. Methods Enzymol 1989; 169: 336-42.
    PubMedGoogle Scholar
  • 27 Rand ML, Perry DW, Packham MA, Gemmell CH, Yeo EL, KinloughRathbone RL. Conditions influencing release of granule contents from human platelets in citrated plasma induced by ADP or the thrombin receptor activating peptide SFLLRN. Am J Hematol 1996; 52: 288-94.
    CrossrefPubMedGoogle Scholar
  • 28 Benistant C, Rubin R. Ethanol inhibits thrombin-induced secretion by human platelets at a site distinct from phospholipase C or protein kinase C. Biochem J 1990; 269: 489-97.
    CrossrefPubMedGoogle Scholar
  • 29 Nishibori M, Cham B, McNicol A, Shalev A, Jain N, Gerrard JM. The protein CD63 is in platelet dense granules, is deficient in a patient with Hermansky-Pudlak syndrome, and appears identical to granulophysin. J Clin Invest 1993; 91: 1775-82.
    CrossrefPubMedGoogle Scholar
  • 30 Fenn CG, Littleton JM. Inhibition of platelet aggregation by ethanol in vitro shows specificity for aggregating agent used and is influenced by platelet lipid composition. Thromb Haemost 1982; 48: 49-53.
    PubMedGoogle Scholar
  • 31 McNicol A, Gerrard JM. Platelet Morphology, Aggregation, and Secretion. In: Adv Molec Cell Biol. The Platelet. Bittar EE, Lapetina EG. eds. Greenwich, Connecticut: JAI Press; 1997. 18 1-29.
    Google Scholar
  • 32 Lemons PP, Chen D, Bernstein AM, Bennett MK, Whiteheart SW. Regulated secretion in platelets: Identification of elements of the platelet exocytosis machinery. Blood 1997; 90: 1490-500.
    PubMedGoogle Scholar
  • 33 Mirlashari MR, Ryningen A, Mikkelsen HM, Fukami MH. Differential secretion of blood platelet storage granules. Platelets 1996; 7: 313-20.
    CrossrefPubMedGoogle Scholar
  • 34 Valdorf-Hansen JF, Zucker MB. 14C-Serotonin release from blood platelets; evidence for more than one mechanism. Am J Physiol 1971; 220: 105-11.
    PubMedGoogle Scholar
  • 35 Packham MA, Kinlough-Rathbone RL, Reimers H-J, Scott S, Mustard JF. Mechanisms of Platelet Aggregation Independent of Adenosine Diphosphate. In: Prostaglandins in Hematology. Silver MJ, Smith JB, Kocsis JJ. eds. New York: Spectrum Publications; 1977: 247-76.
    Google Scholar
  • 36 Stubbs CD, Rubin R. Effect of ethanol on platelet phospholipase A2 . Lipids 1992; 27: 255-60.
    CrossrefPubMedGoogle Scholar