The Effect of Adenosine Monophosphate, Arcaine and Anti-Inflammatory Agents on Thrombosis and Platelet Function in Rabbits

Summary

Thrombi were produced by exposing femoral veins of rabbits to sodium morrhuate followed by partial ligation. The incidence of occlusive thrombosis in 30 min was 95% in animals with or without the infusion of 50-60 ml isotonic saline/kg body weight in 1½–2½ hrs. Comparable infusion of 0.04 M adenosine monophosphate reduced the incidence to 53% (p < 0.01). Arcaine was ineffective even at a toxic concentration. Injection of up to 180 mg phenylbutazone, 25 mg sodium salicylate, or 8.5 mg acetylsalicylic acid (ASA) per kg did not affect the incidence of occlusive thrombosis, but from 15 to 100 mg ASA per kg reduced the incidence to 46% (p < 0.01). Suspensions of connective tissue or collagen particles caused somewhat greater release of platelet-bound ¹⁴C-serotonin in heparinized than in citrated, platelet-rich rabbit plasma, and ASA more readily inhibited connective tissue- or collagen-induced aggregation in the latter. With varying connective tissue concentrations, this drug caused a constant reduction in the amount of ¹⁴C-serotonin released from the platelets in citrated, platelet-rich rabbit plasma, whereas considerably greater inhibition was observed in many samples of citrated, platelet-rich human plasma.

¹⁾ Miss Peterson is now on the staff of the New York Blood Center. Send reprint requests to Dr. Zucker.

• References

• 1 Archer R. K. Haematological Techniques for Use on Animals. Blackwell Scientific Publications; Oxford: 1965
• 2 Baumgartner H. R. Personal communication.
• 3 Baumgartner H. R, Studer A, Reber K. Über Methoden zur Erzeugung experimenteller Thrombosen am Kaninchenohr und die Bedeutung von 5-Hydroxytryptamin bei deren Entstehung. Thrombos. Diathes. haemorrh. (Stuttg.) 9: 485 1963;
• 4 Born G. V. R, Cross M. J. The aggregation of blood platelets. J. Physiol. (Lond.) 168: 178 1963;
• 5 Born G. V. R, Honour A. J, Mitchell J. R. A. Inhibition by adenosine and by 2-chloro-adenosine of the formation and embolization af platelet thrombi. Nature (Lond.) 202: 761 1964;
• 6 Born G. V. R, Philp R. B. Effects of adenosine analogues and of heparin on platelet thrombi in non-lipaemic and lipaemic rats. Brit. J. exp. Path 46: 569 1965;
• 7 Coons A. H. Fluorescent antibody methods. In: General Cytochemical Methods. 399-422 Danielli J. F. Ed. Academic Press; New York: 1958
• 8 Groxton F. E. Elementary Statistics with Applications in Medicine and the Biological Sciences. 276 Dover Publications, Inc.; New York: 1953
• 9 Didisheim P. Inhibition by dipyridamole of arterial thrombosis in rats. Thrombos. Diathes. haemorrh. (Stuttg.) 20: 257 1968;
• 10 Evans G, Packham M. A, Nishizawa E. E, Mustard J. F, Murphy E. A. The effect of acetylsalicylic acid on platelet function. J. exp. Med 128: 877 1968;
• 11 Hampton J. R. The study of platelet behaviour and its relevance to thrombosis. J. athero-scler. Res 7: 729 1967;
• 12 Haslam R. J. Mechanisms of blood platelet aggregation. In: Physiology of Hemostasis and Thrombosis. 88-112 Johnson S. A, and Seegers W. H. Eds. Charles C. Thomas; Springfield, Ill.: 1967
• 13 Henry R. L. Methods for inducing experimental thrombosis: an annotated bibliography. Angiology 13: 554 1962;
• 14 Honour A. J, Mitchell J. R. A. Platelet clumping in injured vessels. Brit. J. exp. Path 45: 75 1964;
• 15 Hovig T. Release of a platelet-aggregating substance (adenosine diphosphate) from rabbit blood platelets induced by saline “extract” of tendons. Thrombos. Diathes. haemorrh. (Stuttg.) 5: 264 1963;
• 16 Jerushalmy Z, Zucker M. B. Some effects of fibrinogen degradation products (FDP) on blood platelets. Thrombos. Diathes. haemorrh. (Stuttg.) 15: 413 1966;
• 17 Leonards J. R. Presence of acetylsalicylic acid in plasma following oral ingestion of aspirin. Proc. Soc. exp. Biol. (N. Y.) 110: 304 1962;
• 18 Macmillan D. C. Secondary clumping effect in human citrated platelet-rich plasma produced by adenosine diphosphate and adrenaline. Nature (Lond.) 211: 140 1966;
• 19 Mills D. C. B, Robb I. A, Roberts G. C. K. The release of nucleotides, 5-hydroxytryptamine and enzymes from human blood platelets during aggregation. J. Physiol. (Lond.) 195: 715 1968;
• 20 Mills D. C. B, Thomas D. P. Blood platelet nucleotides in man and other species. Nature (Lond.) 222: 991 1969;
• 21 Niewiarowski S, Thomas D. P. Personal communication.
• 22 O’Brien J. R. Effects of anti-inflammatory agents on platelets. Lancet I: 894 1968;
• 23 Packham M. A, Warrior E. S, Glynn M. F, Senyi A. S, Mustard J. F. Alteration of the response of platelets to surface stimuli by pyrazole compounds. J. exp. Med 126: 171 1967;
• 24 Pederson H. J, Tebo T. H, Johnson S. A. Evidence of hemolysis in the initiation of hemostasis. Amer. J. clin. Path 48: 62 1967;
• 25 Philp R. B, Lemieux V. Comparison of some effects of dipyridamole and adenosine on thrombus formation, platelet adhesiveness and blood pressure in rabbits and rats. Nature (Lond.) 218: 1072 1968;
• 26 Skoza L, Zucker M. B, Jerushalmy Z, Grant R. Kinetic studies of platelet aggregation induced by adenosine diphosphate and its inhibition by chelating agents, guanidino compounds and adenosine. Thrombos. Diathes. haemorrh. (Stuttg.) 12: 714 1967;
• 27 Spaet T. H, Zucker M. B. Mechanism of platelet plug formation and role of adenosine diphosphate. Amer. J. Physiol 206: 1267 1964;
• 28 Valdorf-Hansen J. F, Zucker M. B. ¹⁴C-serotonin release from blood platelets; evidence for more than one mechanism. (Submitted for publication.)
• 29 Zucker M. B. ADP and collagen-induced platelet aggregation in vivo and in vitro. Thrombos. Diathes. haemorrh. (Stuttg.) Suppl 26: 175 1967;
• 30 Zucker M. B, Peterson J. Inhibition of adenosine diphosphate-induced aggregation and other platelet functions by acetylsalicylic acid ingestion. Proc. Soc. exp. Biol 127: 547 1968;
• 31 Zucker M. B, Peterson J, Grant R. A. Inhibition of some functions of platelets in human citrated platelet-rich plasma by acetylsalicylic acid. Fed. Proc. (abstract) Vol. 28 No. [phone];