Thromb Haemost 1985; 53(03): 306-311
DOI: 10.1055/s-0038-1661303
Original Article
Schattauer GmbH Stuttgart

Endothelial Cells Produce a Lipoxygenase Derived Chemo-Repellent which Influences Platelet/Endothelial Cell Interactions – Effect of Aspirin and Salicylate[*]

M R Buchanan
The Departments of Pathology and Medicine, McMaster University, Hamilton, Ontario, Canada
,
R W Butt
The Departments of Pathology and Medicine, McMaster University, Hamilton, Ontario, Canada
,
Z Magas
The Departments of Pathology and Medicine, McMaster University, Hamilton, Ontario, Canada
,
J Van Ryn
The Departments of Pathology and Medicine, McMaster University, Hamilton, Ontario, Canada
,
J Hirsh
The Departments of Pathology and Medicine, McMaster University, Hamilton, Ontario, Canada
,
D J Nazir
The Departments of Pathology and Medicine, McMaster University, Hamilton, Ontario, Canada
› Author Affiliations
Further Information

Publication History

Received 24 October 1984

Accepted 12 February 1985

Publication Date:
18 July 2018 (online)

Summary

We performed experiments to determine whether endothelial cells synthesize phospholipid metabolites via the lipoxygenase pathway and whether these metabolites influence platelet/vessel wall interactions. Monolayers of cultured human endothelial cells were incubated with 14C-arachidonic acid and their cyclo-oxygenase and lipoxygenase metabolites were extracted and identified by radioimmunoassay, thin layer chromatography and high performance liquid chromatography. We found that in addition to the membrane-associated production of PGI2, endothelial cells synthesized a cytosol-associated metabolite, LOX, which was presumably derived through the lipoxygenase pathway. Inhibition of LOX was associated with an increase in PGI2 production and inhibition of PGI2 with an increase in LOX production. Under either condition, platelet adhesion to cultured endothelial cells was significantly decreased. In contrast, when both PGI2 and LOX production were inhibited, platelet adhesion to endothelial cells was enhanced. Furthermore, when LOX was bound to a thrombogenic surface, platelet adhesion was significantly decreased whereas when arachidonic acid or 12-HETE was bound to the surface, platelet adhesion was increased. We conclude that endothelial cells produce not only a cyclo-oxygenase metabolite, but also a lipoxygenase metabolite, both of which influence platelet/endothelial cell interactions.

Presented, in part, in abstract form at the IXth International Congress Society on Thrombosis and Haemostasis in Stockholm, Sweden, July, 1983.


 
  • References

  • 1 Hamberg M, Samuelsson B. Prostaglandin endoperoxidases. Novel transformations of arachidonic acid in human platelets Proc Natl Acad Sci USA 1974; 71: 3400-3404
  • 2 Hammarstrom S, Falardeau P. Resolution of prostaglandin endoperoxide synthetase and thromboxane synthetase of human platelets. Proc Natl Acad Sci USA 1977; 74: 3691-3695
  • 3 Moncada S, Vane JR. Arachidonic acid metabolites and the interactions between platelets and blood-vessel walls. N Engl J Med 1979; 300: 1142-1147
  • 4 Moncada S, Herman AG, Higgs EA, Vane JR. Differential formation of prostacyclin (PGX or PGI2) by layers of the arterial wall. An explanation for the anti-thrombotic properties of vascular endothelium Thromb Res 1977; 11: 323-344
  • 5 Buchanan MR, Dejana E, Gent M, Mustard JF, Hirsh J. Enhanced platelet accumulation onto injured carotid arteries in rabbits following aspirin treatment. J Clin Invest 1981; 67: 503-508
  • 6 Kelton JG, Hirsh J, Carter CJ, Buchanan MR. Thrombogenic effect of high-dose aspirin in rabbits. Relationship to inhibition of vessel wall synthesis of prostaglandin I2-like activity J Clin Invest 1978; 62: 892-895
  • 7 Buchanan MR, Hirsh J. The effect of aspirin and salicylate on platelet/vessel wall interactions in rabbits. Atherosclerosis 1984; 4: 403-406
  • 8 Buchanan MR, Butt RW, Nazir D, Turpie AG G, Hirsh J. The regulation of platelet/endothelial cell adhesion by two arachidonic acid metabolites. Thromb Haemostas 1983; 50: 190 (Abstr.)
  • 9 Bailey JM, Bryant RW, Feinmark SJ, Makheja AN. Differential separation of thromboxanes from prostaglandins by one and twodimensional thin layer chromatography. Prostaglandins 1977; 13: 479-492
  • 10 Buchanan MR, Blajchman MA, Dejana E, Mustard JF, Senyi AF, Hirsh J. Shortening of the bleeding time in thrombocytopenic rabbits after exposure of jugular vein to high aspirin concentrations. Prostaglandins Med 1979; 13: 333-342
  • 11 Kinlough-Rathbone RL, Mustard JF, Packham MA, Perry DW, Reimers H-J, Cazenave J-P. Properties of washed human platelets. Thromb Haemostas 1977; 37: 291-308
  • 12 Gimbrone MA, Buchanan MR. Interactions of platelets and leukocytes with vascular endothelium: In vitro studies. In Endothelium Fishmann AP. (Ed) Ann NY Acad Sci 1983; 401: 171-183
  • 13 Buchanan MR, Butt RW, Markham B, Hirsh J. Effects of aspirin and salicylate on platelet function. Thromb Haemostas 1983; 50: 101
  • 14 Gimbrone MA, Shefton ES, Cruise SA. Isolation and primary culture of endothelial cells from human umbilical vessels. TCA Manual 1978; 4: 813-817
  • 15 Buchanan MR, Rischke JA, Butt RW, Turpie AG G, Hirsh J, Rosenfeld J. The sex-related differences in aspirin pharmacokinetics in rabbits and man and its relationship to antiplatelet effects. Thromb Res 1983; 29: 125-139
  • 16 Schafer AI. Deficiency of platelet lipoxygenase activity in myeloproliferative disorders. N Engl J Med 1982; 306: 381-386
  • 17 Roth GJ, Majerus PW. Acetylation of a particulate fraction protein. J Clin Invest 1975; 56: 624-632
  • 18 Siegel MI, McConnell RT, Cuatrecasas P. Aspirin-like drugs interfere with arachidonate metabolism by inhibition of the 12-hydroperoxy-5,8,10,14-eicosatetraenoic acid peroxidase activity of the lipoxygenase pathway. Proc Natl Acad Sci USA 1979; 76: 3774-3778
  • 19 Wilhelm TE, Sankarappa SK, Van Rollins M, Sprecher H. Selective inhibitors of platelet lipoxygenase: 4,7,10,13-icosatetraynoic acid and 5,8,11,14-henicosatetraynoic acid. Prostaglandins 1981; 21: 323-332
  • 20 Dunnett CW. A multiple comparison procedure for comparing several treatments with a control. Amer Statist Assoc J 1955; 50: 1096-1106
  • 21 Dunnett CW. New tables for multiple comparisons with a control. Biometrics 1964; 20: 482-491
  • 22 Sawyer PN, Srinivasan S. The role of electrochemical surface properties in thrombosis at vascular interfaces: cumulative experience of studies in animals and man. Bull NY Acad Med 1972; 48: 235-256
  • 23 Hammarstrom S. Leukotrienes: cellular regulators formed by conjugation of polyunsaturated fatty acids with glutathione. In Reviews in Biochemical Toxicology. Hodgson E, Benel JR, Philpot RM. (Eds) Elsevier Science Publishing Co Inc: New York, New York: 1983. p 173
  • 24 Nugteren DH. Arachidonate lipoxygenase in blood platelets. Biochim Biophys Acta 1975; 380: 299-307
  • 25 Smith JB. Prostaglandins and platelet aggregation. Acta Med Scand 1981; (Suppl) (Suppl. 210) 651: 91-99
  • 26 Malmsten CL. The effects of some non-steroidal anti-inflammatory drugs upon LTB4 formation and granulocyte function in vitro. In Prostaglandins and Leukotrienes ’84. Abst Vol. Bailey JM. (Ed) George Washington Univ Press; Washington D C: 1984. p 15
  • 27 Gorman RR. Modulation of human platelet function by prostacyclin and thromboxane A2 . Fed Proc 1979; 38: 83-88