The Influence of Ascorbic Acid on Platelet Structure and Function

Dale H Cowan; Richard C Graham Jr.; Patricia Shook; Ronda Griffin

doi:10.1055/s-0038-1651445

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1975; 34(01): 050-062
DOI: 10.1055/s-0038-1651445

Original Article

Schattauer GmbH

The Influence of Ascorbic Acid on Platelet Structure and Function

Dale H Cowan

¹Department of Medicine, Case Western Reserve University School of Medicine at Cleveland Metropolitan General Hospital and University Hospitals, Cleveland, Ohio, U.S.A.

,

Richard C Graham Jr.

¹Department of Medicine, Case Western Reserve University School of Medicine at Cleveland Metropolitan General Hospital and University Hospitals, Cleveland, Ohio, U.S.A.

,

Patricia Shook

¹Department of Medicine, Case Western Reserve University School of Medicine at Cleveland Metropolitan General Hospital and University Hospitals, Cleveland, Ohio, U.S.A.

,

Ronda Griffin

¹Department of Medicine, Case Western Reserve University School of Medicine at Cleveland Metropolitan General Hospital and University Hospitals, Cleveland, Ohio, U.S.A.

› Author Affiliations

Abstract

Summary

To determine the effect on platelet behavior of transient exposure of platelets to ascorbic acid, studies of platelet function and ultrastructure were done before exposure to ascorbic acid at pH 6.5, during exposure to pH 6.5, and after restoration of pH to pre-acidifìcation levels. The effect of ascorbic acid (A. A.) was compared to that of HCl and citric acid (C. A.). ADP- and collagen-induced aggregation of normal platelets were significantly impaired by both A. A. and C. A. but were less affected by HCl. The release of ¹⁴C-serotonin was significantly reduced by each agent. The ultra-structure of normal platelets brought to pH 6.5 by A.A. was normal. After neutralization, there was marked dilatation of the open channel system and loss of the disc shape. When platelets were brought to pH 6.5 by A. A., then neutralized, the aggregates which formed after stimulation by ADP or collagen were smaller than normal, the platelets were less closely approximated, and degranulation was less complete. The data show that exposure of platelets to ascorbic acid for short intervals impairs their function when measured after restoration of pH to levels compatible with maximal responses. Platelet survival studies using autologous platelets labelled with ⁵¹Cr in the presence or absence of ascorbic acid showed that the recovery of normal platelets was unaffected by ascorbic acid, whereas recovery of platelets from patients with idiopathic thrombocytopenic purpura, idiopathic thrombocythemia, and alcohol-related thrombocytopenia was markedly reduced. The injury resulting from the use of ascorbic acid in preparing platelets for studies of platelet survival in patients with disorders affecting platelets may impair the recovery of the cells, resulting in artifactual changes in the survival studies.

Full Text

References

References
1 Aledort L. M, Troup S. B, and Weed R. I. 1968; Inhibition of sulfhydryl-dependent platelet functions by penetrating and non-penetrating analogues of parachloromercuri-benzene. Blood 31: 47.
2 Aster R. H. 1971. Factors affecting the kinetics of isotopically labelled platelets. In: Paulus J. M. (Ed.). Platelet Kinetics, Radioisotopic, Cytological, Mathemacial, and Clinical Aspects. North Holland-American Elsevier; Amsterdam, London, New York: 3.
3 Aster R. H, and Jandl J. H. 1964; Platelet sequestration in man. I. Methods. Journal of Clinical Investigation 43: 843.
4 Born G. V. R, and Cross M. J. 1963; The aggregation of blood platelets. Journal of Physiology 168: 178.
5 Brecher G, Schneiderman M, and Cronkite E. P. 1953; The reproducibility and constancy of the platelet count. American Journal of Clinical Pathology 23: 15.
6 Clancy R, Jenkins E, and Firkin B. 1972; Qualitative platelet abnormalities in idiopathic thrombocytopenic purpura. New England Journal of Medicine 286: 622.
7 Cowan D. H. 1973; Thrombokinetic studies in alcohol-related thrombocytopenia. The Journal of Laboratory and Clinical Medicine 81: 64.
8 Fantl P, Ebbels L, and Nelson J. F. 1951; The presence of thiol groups in thrombocytes and their significance in the contraction of fibrin gel. British Journal of Experimental Pathology 32: 538.
9 Flatow Jr. F. A, and Freireich E. J. 1966; The increased effectiveness of platelet concentrates prepared in acidified plasma. Blood 27: 449.
10 Haut M. J, and Cowan D. H. 1974; The effect of ethanol on the hemostatic properties of human blood platelets. American Journal of Medicine 56: 22.
11 Kattlove H. E, and Spaet II T. 1970; The effect of chromium on platelet function in vitro. Blood 35: 659.
12 Kikugawa K, Iizuka K, and Ichino M. 1972; Effect of plasma pH on stability and capacity of aggregation of platelets. Chemical Pharmacological Bulletin 20: 1569.
13 Lamberth E. L, Warriner III R. A, and Batchelor E. D. 1974; Effect of metabolic acidosis and alkalosis on human platelet aggregation induced by epinephrine and ADP. Proceedings of the Society for Experimental Biology and Medicine 145: 743.
15 McClure P. D, Ingram G. I. C, Stacey R. S, Glass U. H, and Matchett M. O. 1966; Platelet function tests in thrombocythaemia and thrombocytosis. British Journal of Haematology 12: 478.
16 Robinson Jr. C. W, Mason R. G, and Wagner R. H. 1963; Effect of sulfhydryl inhibitors on platelet agglutinability. Proceedings of the Society for Experimental Biology and Medicine 113: 857.
17 Rodman N. F, and Penick G. D. 1972; The effect of pH on platelet aggregation responses. Blood 40: 953 (abstract).
18 Rogers A. B. 1972; The effect of pH on human platelet aggregation induced by epinephrine and ADP. Proceedings of the Society for Experimental Biology and Medicine 139: 1100.
19 Sherman R. W, Mason R. G, Wagner R. H, and Brinkhotjs K. M. 1961; Studies on thrombin-induced platelet agglutination. Journal of Experimental Medicine 114: 905.
20 Spaet T. H, Lejnieks I, Gaynor E, and Goldstein M. C. 1969; Defective platelets in essential thrombocythemia. Archives of Internal Medicine 124: 135.
21 Valdorf-Hansen J. F, and Zucker M. B. 1971; Effect of temperature and inhibitors on serotonin-¹⁴C release from human platelets. American Journal of Physiology 220: 105.
22 Walsh P. N. 1972; Albumin density gradient separation and washing of platelets and the study of platelet coagulant activities. British Journal of Haematology 22: 205.
23 White J. G. 1968; Fine structural alterations induced in platelets by adenosine diphosphate. Blood 31: 604.