Clotting Activity of Platelet Phospholipids in Rat and Man

Summary

The clotting activity of man and rat platelet phospholipid fractions separated by bi-dimensional TLC, resuspended in Tyrode by sonication, was studied in the recalcification (manual) and in the Stypven (recalcification plus Russell’s viper venom) clotting time (determined in a coagulometer). Phosphatidyl serine was the most active fraction to shorten the two clotting tests utilized, in both rat and man, but it was much more effective in the Stypven time. The phosphatidyl ethanolamine was the second most active fraction, in the Stypven time; this fraction was almost as active as phosphatidyl serine in both animal species. The other fractions studied (phosphatidyl inositol, phosphatidyl choline and sphingomyelin) were sligthly active or not active, depending on the experimental conditions.

The clotting activity of platelet phosphatidyl serine from rat, at concentrations corresponding to platelet counts from 1 to 10 ( X10⁵), was much smaller than this of the disrupted (sonicated) platelets from which it originated. However, the clotting activity of sonicated platelets could be completely reproduced, either at each concentration studied (Stypven time) or at a concentration corresponding to lOxlO⁵ platelets (recalcification time), by adding to phosphatidyl serine the other four phospholipid fractions (phosphatidyl inositol, phosphatidyl ethanolamine, phosphatidyl choline, sphingomyelin) dispersed in a homogeneous way by sonication.

The feeding of a butter-rich diet to rats considerably increased the activity of each of the platelet phospholipid fractions in the two clotting tests carried out.

^* Attache de Recherche, D.G.R.S.T. Contract N° 73- 7- 1050

^** Maitre de Recherch e, I.N.S.E.R.M.

• References

• 1 Folch J, Lees M, Sloane G. HStanely. 1957; A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biochemistry 226: 497.
• 2 Gautheron P, Renaud S. 1972; Hyperlipemia induced hypercoagulable state in rat. Role of an increased activity of platelet phosphatidyl serine in response to certain dietary fatty acids. Thrombosis Research 01: 353.
• 3 Marcus A. J. 1967. The role of lipids in blood coagulation. In: Advances in Lipid Research. Academic Press; New York.:
• 4 Marcus A. J, Ullman H. L, Safier L. B, Ballard H. S. 1962; Platelet phospholipids. Their fatty acid and aldehyde composition and activity in different clotting systems. Journal of Clinical Investigation 41: 2198.
• 5 Renaud S, Gautheron P. 1974. b Dietary fats and experimental (cardiac and venous) thrombosis. In: Dietary fats and Thrombosis. S. Karger; Basle: 53.
• 6 Renaud S, Gautheron P, Arbogast R, Dumont E. 1974; Platelet factor 3 activity and platelet aggregation in patients submitted to coronarography. Scandinavian Journal of Haematology 12: 85.
• 7 Renaud S, Gautheron P, Rosenstein H. 1973; Platelet factor 3 activity in washed platelets. Thrombosis Diathesis Hæmorrhogica 30: 557.
• 8 Renaud S, Gautheron P. 1974 a Influence of dietary fats on atherosclerosis, coagulation and platelet phospholipids in rabbit. Atherosclerosis. (In Press)
• 9 Renaud S, Lecompte F. 1970; Hypercoagulability induced by hyperlipemia in rat, rabbit and man. Role of platelet factor 3. Circulation Research 27: 1003.
• 10 Troup S. B, Reed C. F, Marinetti G. V, Swisher S. N. 1960; Thromboplastic factors in platelets and red blood cells: observations on their chemical nature and function in in vitro coagulation. Journal of Clinical Investigation 39: 342.
• 11 Woodside E. E, Therriault D. G, Kocholaty W. 1964; Lipids of human platelets and their action on the blood coagulation process. Blood 24: 76.