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. 1978 Oct;62(4):847–856. doi: 10.1172/JCI109197

Effect of Aspirin on Thrombin-Induced Adherence of Platelets to Cultured Cells from the Blood Vessel Wall

Robert L Czervionke 1, John C Hoak 1, Glenna L Fry 1
PMCID: PMC371837  PMID: 701483

Abstract

An in vitro method was used to detect adherence of 51Cr-labeled platelets to monolayers of cultured human endothelial, fibroblast, and smooth muscle cells. Washed platelets did not adhere to untreated or aspirin-treated endothelial monolayers in the absence of thrombin. In contrast, thrombin-induced platelet aggregates adhered to all of the monolayers but adherence to endothelium was significantly less than to the other cells. Additional evidence for adherence of platelets to the endothelium was provided by scanning and transmission electron microscopy. Thrombin-induced platelet adherence to endothelium was inhibited by hirudin. Platelet adherence induced by thrombin was enhanced significantly by treatment of the endothelial monolayer with 1—2 mM aspirin. This increase in adherence was seen even when aspirin-treated platelets were used; adherence values approached those seen with fibroblasts and smooth muscle cells. An aspirin concentration of 0.1 mM was sufficient to block thrombin-induced malonaldehyde production in platelets but it did not interfere with the inhibitory effect of the endothelium against platelet adherence. The effect of aspirin on the endothelium was temporary and inhibitory activity of the endothelium was restored 1 h after aspirin had been removed from the incubation system. The ability of thrombin to cause adherence of platelets to undamaged endothelium, and the potential for aspirin to enhance this adherence have implications for mechanisms which operate in platelet interaction with the blood vessel wall.

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Selected References

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  1. ASTER R. H., JANDL J. H. PLATELET SEQUESTRATION IN MAN. I. METHODS. J Clin Invest. 1964 May;43:843–855. doi: 10.1172/JCI104970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baenziger N. L., Dillender M. J., Majerus P. W. Cultured human skin fibroblasts and arterial cells produce a labile platelet-inhibitory prostaglandin. Biochem Biophys Res Commun. 1977 Sep 9;78(1):294–301. doi: 10.1016/0006-291x(77)91253-0. [DOI] [PubMed] [Google Scholar]
  3. Baumgartner H. R., Stemerman M. B., Spaet T. H. Adhesion of blood platelets to subendothelial surface: distinct from adhesion to collagen. Experientia. 1971 Mar 15;27(3):283–285. doi: 10.1007/BF02138148. [DOI] [PubMed] [Google Scholar]
  4. Baumgartner H. R. The role of blood flow in platelet adhesion, fibrin deposition, and formation of mural thrombi. Microvasc Res. 1973 Mar;5(2):167–179. doi: 10.1016/0026-2862(73)90069-1. [DOI] [PubMed] [Google Scholar]
  5. Buonassisi V., Root M. Enzymatic degradation of heparin-related mucopolysaccharides from the surface of endothelial cell cultures. Biochim Biophys Acta. 1975 Mar 14;385(1):1–10. doi: 10.1016/0304-4165(75)90067-7. [DOI] [PubMed] [Google Scholar]
  6. Burch J. W., Stanford N., Majerus P. W. Inhibition of platelet prostaglandin synthetase by oral aspirin. J Clin Invest. 1978 Feb;61(2):314–319. doi: 10.1172/JCI108941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cazenave J. P., Packham M. A., Guccione M. A., Mustard J. F. Inhibition of platelet adherence to damaged surface of rabbit aorta. J Lab Clin Med. 1975 Oct;86(4):551–563. [PubMed] [Google Scholar]
  8. Dosne A. M., Drouet L., Dassin E. Usefulness of 51chromium-platelet labelling for the measurement of platelet deposition on subendothelium. Microvasc Res. 1976 Jan;11(1):111–114. doi: 10.1016/0026-2862(76)90082-0. [DOI] [PubMed] [Google Scholar]
  9. Gimbrone M. A., Jr, Cotran R. S. Human vascular smooth muscle in culture. Growth and ultrastructure. Lab Invest. 1975 Jul;33(1):16–27. [PubMed] [Google Scholar]
  10. Jaffe E. A., Nachman R. L., Becker C. G., Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. doi: 10.1172/JCI107470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kelley R. O., Dekker R. A., Bluemink J. G. Ligand-mediated osmium binding: its application in coating biological specimens for scanning electron microscopy. J Ultrastruct Res. 1973 Nov;45(3):254–258. doi: 10.1016/s0022-5320(73)80051-6. [DOI] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Lange W. E., Bell S. A. Fluorometric determination of acetylsalicylic acid and salicylic acid in blood. J Pharm Sci. 1966 Apr;55(4):386–389. doi: 10.1002/jps.2600550407. [DOI] [PubMed] [Google Scholar]
  14. Lough J., Moore S. Endothelial injury induced by thrombin or thrombi. Lab Invest. 1975 Aug;33(2):130–135. [PubMed] [Google Scholar]
  15. McDonald R. I., Shepro D., Rosenthal M., Booyse F. M. Properties of cultured endothelial cells. Ser Haematol. 1973;6(4):469–478. [PubMed] [Google Scholar]
  16. Moncada S., Gryglewski R., Bunting S., Vane J. R. An enzyme isolated from arteries transforms prostaglandin endoperoxides to an unstable substance that inhibits platelet aggregation. Nature. 1976 Oct 21;263(5579):663–665. doi: 10.1038/263663a0. [DOI] [PubMed] [Google Scholar]
  17. Moncada S., Higgs E. A., Vane J. R. Human arterial and venous tissues generate prostacyclin (prostaglandin x), a potent inhibitor of platelet aggregation. Lancet. 1977 Jan 1;1(8001):18–20. doi: 10.1016/s0140-6736(77)91655-5. [DOI] [PubMed] [Google Scholar]
  18. Mustard J. F., Perry D. W., Ardlie N. G., Packham M. A. Preparation of suspensions of washed platelets from humans. Br J Haematol. 1972 Feb;22(2):193–204. doi: 10.1111/j.1365-2141.1972.tb08800.x. [DOI] [PubMed] [Google Scholar]
  19. O'Brien J. R. Effects of salicylates on human platelets. Lancet. 1968 Apr 13;1(7546):779–783. doi: 10.1016/s0140-6736(68)92228-9. [DOI] [PubMed] [Google Scholar]
  20. Okuma M., Steiner M., Baldini G. Studies on lipid peroxides in platelets. II. Effect of aggregating agents and platelet antibody. J Lab Clin Med. 1971 May;77(5):728–742. [PubMed] [Google Scholar]
  21. Rafelson M. E., Jr, Hoveke T. P., Booyse F. M. The molecular biology of platelet-platelet and platelet-endothelial interactions. Ser Haematol. 1973;6(3):367–381. [PubMed] [Google Scholar]
  22. Rome L. H., Lands W. E., Roth G. J., Majerus P. W. Aspirin as a quantitative acetylating reagent for the fatty acid oxygenase that forms prostaglandins. Prostaglandins. 1976 Jan;11(1):23–30. doi: 10.1016/0090-6980(76)90169-6. [DOI] [PubMed] [Google Scholar]
  23. Ross R. The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol. 1971 Jul;50(1):172–186. doi: 10.1083/jcb.50.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roth G. J., Stanford N., Majerus P. W. Acetylation of prostaglandin synthase by aspirin. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3073–3076. doi: 10.1073/pnas.72.8.3073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rowland M., Riegelman S., Harris P. A., Sholkoff S. D. Absorption kinetics of aspirin in man following oral administration of an aqueous solution. J Pharm Sci. 1972 Mar;61(3):379–385. doi: 10.1002/jps.2600610312. [DOI] [PubMed] [Google Scholar]
  26. Saba S. R., Mason R. G. Studies of an activity from endothelial cells that inhibits platelet aggregation, serotonin release, and clot retraction. Thromb Res. 1974 Dec;5(6):747–757. doi: 10.1016/0049-3848(74)90118-2. [DOI] [PubMed] [Google Scholar]
  27. Saba S. R., Zucker W. H., Mason R. G. Some properties of endothelial cells isolated from human umbilical cord vein. Ser Haematol. 1973;6(4):456–468. [PubMed] [Google Scholar]
  28. Smith J. B., Ingerman C. M., Silver M. J. Malondialdehyde formation as an indicator of prostaglandin production by human platelets. J Lab Clin Med. 1976 Jul;88(1):167–172. [PubMed] [Google Scholar]
  29. Stemerman M. B., Baumgartner H. R., Spaet T. H. The subendothelial microfibril and platelet adhesion. Lab Invest. 1971 Mar;24(3):179–186. [PubMed] [Google Scholar]
  30. Stemerman M. B., Spaet T. H. The subendothelium and thrombogenesis. Bull N Y Acad Med. 1972 Feb;48(2):289–301. [PMC free article] [PubMed] [Google Scholar]
  31. Stuart M. J., Murphy S., Oski F. A. A simple nonradioisotope technic for the determination of platelet life-span. N Engl J Med. 1975 Jun 19;292(25):1310–1313. doi: 10.1056/NEJM197506192922502. [DOI] [PubMed] [Google Scholar]
  32. Villa S., de Gaetano G. Prostacyclin-like activity in rat vascular tissues. Fast, long-lasting inhibition by treatment with lysine acetylsalicylate. Prostaglandins. 1977;14(6):1117–1124. doi: 10.1016/0090-6980(77)90289-1. [DOI] [PubMed] [Google Scholar]
  33. Wasteson A., Glimelius B., Busch C., Westermark H., Heldin C. H., Norling B. Effect of a platelet endoglycosidase on cell surface associated heparan sulphate of human culturei endothelial and glial cells. Thromb Res. 1977 Sep;11(3):309–321. doi: 10.1016/0049-3848(77)90184-0. [DOI] [PubMed] [Google Scholar]
  34. Weiss H. J., Aledort L. M., Kochwa S. The effect of salicylates on the hemostatic properties of platelets in man. J Clin Invest. 1968 Sep;47(9):2169–2180. doi: 10.1172/JCI105903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Weksler B. B., Marcus A. J., Jaffe E. A. Synthesis of prostaglandin I2 (prostacyclin) by cultured human and bovine endothelial cells. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3922–3926. doi: 10.1073/pnas.74.9.3922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Winkelhake J. L., Nicolson G. L. Determination of adhesive properties of variant metastatic melanoma cells to BALB/3T3 cells and their virus-transformed derivatives by a monolayer attachment assay. J Natl Cancer Inst. 1976 Feb;56(2):285–291. doi: 10.1093/jnci/56.2.285. [DOI] [PubMed] [Google Scholar]

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