Download PDF
Thromb Haemost 1984; 52(02): 205-209
DOI: 10.1055/s-0038-1661173
Original Article
Schattauer GmbH Stuttgart

Effect of Adenyl-Cyclase Activators, Phosphodiesterase Inhibitors and Pyridoxal-5-Phosphate on Platelet Aggregation and Adenosine-3’-5’-Cyclic Monophosphate Accumulation

M Zahavi
1   The Department of Medicine, Day Clinic and the Vascular Laboratory, Tel-Aviv Medical Center, Tel- Aviv, Israel
,
J Zahavi
1   The Department of Medicine, Day Clinic and the Vascular Laboratory, Tel-Aviv Medical Center, Tel- Aviv, Israel
,
V V Kakkar
2   The Thrombosis Research Unit, King’s College Hospital and Medical School, London, U. K.
› Author Affiliations
Further Information

Publication History

Received 16 March 1984

Accepted 14 June 1984

Publication Date:
19 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

The effect of prostaglandin E1 (PGE1) and compound BW245C, adenyl-cyclase activators, theophylline, papaverine and dipyridamole, phosphodiesterase (PDE) inhibitors and pyridoxal- 5-phosphate (PALP) on inhibition of platelet aggregation (PA) and platelet c-AMP accumulation was determined in human platelet-rich plasma (PRP) and washed platelets. PGE1 at 280 nM and BW245C at 7.7 nM induced a significant PA inhibition in PRP and washed platelets (though less pronounced by PGE1) concomitant to a very large increase (8-13-fold) in platelet cAMP level both in PRP and washed platelets. At comparable PA inhibition, c-AMP level was not significantly changed by PALP and only moderately (but significantly) increased (2-4.6-fold) by PDE inhibitors. PALP or theophylline potentiated both PGE1 induced platelet c-AMP accumulation and PA inhibition. Yet PALP potentiated theophylline-induced PA inhibition without affecting platelet c-AMP level.

Our results indicate that 2 c-AMP pools are presumably present in the platelets, since at comparable c-AMP accumulation PDE inhibitors or BW245C were more effective than PGE1 in PA inhibition in PRP. Morever, this pattern was more pronounced in washed platelets and was also found in the presence of thrombin and adenosine diphosphate which induced a decrease in platelet c-AMP level. The effect of PALP on PA inhibition is presumably mediated by 2 mechanisms: a) a direct effect on the platelet membrane independent from the c-AMP system, b) In the presence of PGE1, the increment in PA inhibition, is through further indirect activation of the adenyl-cyclase. The mechanism of PA inhibition by BW245C in intact platelets, is most probably through activation of adenyl-cyclase. This study emphasize the importance of c-AMP accumulation in the inhibition of PA.

Keywords

c-AMP - Platelet aggregation inhibition - Pyridoxal-5-phos-phate - Compound BW245C
 

  • References

  • 1 Haslam RJ, Davidson MM L, Davies T, Lynham JA, McLenaghan MD. Regulation of blood platelet function by cyclic nucleotides. In: Advances in Cyclic Nucleotide Research. Hamet P, Sands H. (eds) Raven Press; New York: 1978. 9 533-552
    Google Scholar
  • 2 Steer ML, Salzman EW. Cyclic nucleotides in haemostasis and thrombosis. In: Advances in Cyclic Nucleotide Research. Hamet P, Sands H. (eds) Raven Press; New York: 1980. 12 71-92
    Google Scholar
  • 3 Ball G, Breveton GG, Fulwood M, Ireland DM, Yates P. Effect of prostaglandin E1alone and in combination with theophylline or aspirin on collagen-induced platelet aggregation and on platelet nucleotides adenosine-3’-5’-cyclic monophosphate. Biochem J 1970; 120: 709-718
    CrossrefPubMedGoogle Scholar
  • 4 Mills DC B, Smith JB. The influence on platelet aggregation of drugs that affect the accumulation of adenosine-3’-5’-cyclic monophosphate in platelets. Biochem J 1971; 121: 185-196
    CrossrefPubMedGoogle Scholar
  • 5 Zahavi M, Zahavi J, Kakkar VV. Modulation of platelet function by adenosine-3’-5’-cyclic monophosphate. Thromb Haemostas 1983; 50: 450 (Abstr.)
    PubMedGoogle Scholar
  • 6 Subbarrao K, Kakkar VV, Ganguly P. Binding of pyridoxal-5- phosphate to human platelets: its effect on platelet function. Thromb Res 1978; 13: 1017-1029
    CrossrefPubMedGoogle Scholar
  • 7 Subbarao K, Kuchibholta J, Kakkar VV. Pyridoxal-5-phosphate - a new physiological inhibitor of blood coagulation and platelet function. Biochem Pharmacol 1979; 28: 531-534
    CrossrefPubMedGoogle Scholar
  • 8 Lam SC T, Harfenist MA, Packham MA, Mustard JF. Investigation of possible mechanisms of pyridoxal-5-phosphate inhibition of platelet reactions. Thromb Res 1980; 20: 633-645
    CrossrefPubMedGoogle Scholar
  • 9 Celia G, Zahavi J, deHaas HA, Kakkar VV. β-thromboglobulin, platelet production time and platelet function in vascular disease. Br J Haematol 1979; 43: 127-136
    CrossrefPubMedGoogle Scholar
  • 10 Zahavi M, Zahavi J, Kakkar VV. The effect of pyridoxal-5-phosphate on the inhibition of platelet aggregation and adenosine-3’-5’-cyclic monophosphate accumulation in human platelets. Life Sciences. 1984 (in press)
    PubMedGoogle Scholar
  • 11 Brown BL, Albano JD M, Ekins RP, Scherzi AM. A simple and sensitive saturation assay method for the measurement of adenosine- 3’-5’-cyclic monophosphate. Biochem J 1971; 121: 561-562
    CrossrefPubMedGoogle Scholar
  • 12 Gilman AJ, Murad F. Assay of cyclic nucleotide by receptor protein binding displacement. Methods Enzymol 1974; 38: 49-61
    PubMedGoogle Scholar
  • 13 McDonald JW D, Stuart RK. Regulation of c-AMP levels and aggregation in human platelets by prostaglandin E1 . J Lab Clin Med 1973; 81: 838-849
    PubMedGoogle Scholar
  • 14 Cole B, Robinson GA, Hartman RC. Studies on the role of cyclic AMP in platelet function. Ann NY Acad Sci 1971; 185: 477-487
    CrossrefPubMedGoogle Scholar
  • 15 Haslam RJ, Rosson GM. Effects of adenosine on levels of adenosine cyclic-3’-5’-monophosphate in human platelets in relation to adenosine incorporation and platelet aggregation. Mol Pharmacol 1975; 11: 528-544
    PubMedGoogle Scholar
  • 16 Vermylen J, Badenhorst PN, Deckmyn H, Arnout J. Normal mechanisms of platelet. Clin Haematol 1983; 12: 107-151
    PubMedGoogle Scholar
  • 17 Lam SC T, Guccione MA, Packham MA, Mustard JF. Effect of c-AMP phosphodiesterase inhibitors on ADP-induced shape change, c-AMP and nucleoside diphosphokinase activity of rabbit platelets. Thromb Haemostas 1982; 47: 90-95
    PubMedGoogle Scholar
  • 18 Haslam RJ. Role of cyclic nucleotides in platelet function. In: Biochemistry and Pharmacology of Platelets. Ciba Foundation Symposium, 35, Elsevier, North Holland Biomedical Press; Amsterdam: 1975. pp 121-151
    Google Scholar
  • 19 Tsien WH, Saas SP, Sheppard H. The lack of correlation between inhibition of aggregation and c-AMP levels with canine platelets. Thromb Res 1982; 28: 509-519
    CrossrefPubMedGoogle Scholar
  • 20 Buxton IL O, Brunton LL. Compartments of cyclic AMP and protein kinase in mammalian cardiomyocytes. J Biol Chem 1983; 258: 10233-10239
    PubMedGoogle Scholar
  • 21 Sinha AK, Colman RW. Prostaglandin E1inhibits platelet aggregation by a pathway independent of adenosine-3’-5’-cyclic monophosphate. Nature 1978; 200: 202-203
    PubMedGoogle Scholar
  • 22 Sinha AK, Colman RW. Cyclic AMP independent inhibition of platelet aggregation by prostaglandin Ei is mediated by factor Xa. Thromb Res 1983; 30: 553-564
    CrossrefPubMedGoogle Scholar
  • 23 Imakoa T, Lynham JA, Haslam RJ. Purification and characterization of the 47,000 dalton protein phosphorylated during degranulation of human platelets. J Biol Chem 1983; 258: 11404-11414
    PubMedGoogle Scholar
  • 24 Kaeser-Glanzmann R, Gerber E, Lüscher E. Regulation of the intracellular calcium level in human blood platelets: cyclic adenosine- 3’-5’-monophosphate dependent phosphorylation of a 22,000 dalton component in isolated Ca++-accumulating vesicles. Biochim Biophys Acta 1979; 558: 344-347
    CrossrefPubMedGoogle Scholar