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Thromb Haemost 2000; 83(02): 338-344
DOI: 10.1055/s-0037-1613809
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Homocysteine and Oxidized Low Density Lipoprotein Enhance Platelet Adhesion to Endothelial Cells under Flow Conditions: Distinct Mechanisms of Thrombogenic Modulation

R. Dardik
,
D. Varon
,
I. Tamarin
,
A. Zivelin
,
O. Salomon
,
B. Shenkman
,
N. Savion
1   Goldschleger Eye Research Institute, Tel Aviv University, Tel Aviv, Israel
› Author Affiliations
Further Information

Publication History

Received 26 November 1998

Accepted after resubmission 31 August 1999

Publication Date:
11 December 2017 (online)

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Summary

We investigated the effects of two well established risk factors for cardiovascular disease, homocysteine and oxidized low density lipoprotein (ox-LDL), on endothelial cell thrombogenicity. For this purpose we studied platelet adhesion to human endothelial cells (EC) under flow conditions at a shear rate of 350 s−1 following EC treatment with either homocysteine or ox-LDL. Treatment of EC with either homocysteine (1 or 10 mmol/L for 16 h) or ox-LDL (100 µg/ml for 16 h) resulted in a 2-3 fold enhancement in platelet adhesion. The enhancement in platelet adhesion induced by 1 mmol/L homocysteine, but not that induced by 10 mmol/L homocysteine, was absolutely dependent on fibrin formation. Homocysteine treatment has significantly increased the cell surface tissue factor (TF) activity and slightly reduced the expression of the intercellular adhesion molecule I (ICAM-1). In contrast, ox-LDL treatment upregulated ICAM-1 expression and had no significant effect on endothelial TF activity. Neither homocysteine nor Ox-LDL affected surface expression of the αvβ3 integrin. The homocysteine-induced enhancement in platelet adhesion was almost completely abolished by blockade of the EC TF activity by a polyclonal antibody. The enhancing effect of homocysteine was also greatly reduced by inhibition of the EC αvβ3 integrin, but was not affected by blockade of EC ICAM-1. On the other hand, ox-LDL-induced enhancement in platelet – EC adhesion was greatly inhibited by blocking ICAM-1 or αvβ3, but remained unaffected by inhibition of TF activity. Preincubation of platelets with the glycoprotein IIb-IIIa (GPIIb-IIIa) antagonist Reo-Pro has virtually abolished the enhancing effect of both homocysteine and ox-LDL. Our results suggest that homocysteine and ox-LDL might increase endothelial thrombogenicity by distinct mechanisms: homocysteine – by inducing TF activity, and ox-LDL – by upregulating ICAM-1, both of which enhance GPIIb-IIIa/fibrinogen dependent platelet adhesion to EC. The αvβ3 integrin, although not affected by EC stimulation, seems to play a crucial role in platelet-EC interaction regardless of the mechanism of EC perturbation.

Keywords

Homocysteine oxidized LDL - endothelial cells
 

  • References

  • 1 McCully KS. Homocysteine and vascular disease. Nature Medicine 1996; 02: 386-9.
    CrossrefPubMedGoogle Scholar
  • 2 Moncada S, Palmer RMJ, Higgs EA. Prostacyclin and endothelium-derived relaxing factor: biological interactions and significance. In: Thrombosis and Heamostasis 1987. Verstraete M, Vermylen J, Lijnen R, Arnout J. (eds). Leuven: Leuven University Press; 597-618.
    Google Scholar
  • 3 Lentz SR, Sadler JE. Inhibition of thrombomodulin surface expression and protein C activation by the thrombogenic agent homocysteine. J Clin Invest 1991; 88: 1906-14.
    CrossrefPubMedGoogle Scholar
  • 4 Rodgers GM, Conn MT. Homocysteine, am atherogenic stimulus, reduces protein C activation by arterial and venous endothelial cells. Blood 1990; 75: 895-901.
    PubMedGoogle Scholar
  • 5 Ratnoff OD. Activation of Hageman factor by L-homocystine. Science 1968; 162: 1007-9.
    PubMedGoogle Scholar
  • 6 Rodgers GM, Kane WH. Activation of endogenous factor V by a homocysteine-induced vascular endothelial cell activator. J Clin Invest 1986; 77: 1909-16.
    CrossrefPubMedGoogle Scholar
  • 7 Fryer RH, Wilson BD, Gubler DB, Fitzgerald LA, Rodgers GM. Homocysteine, a risk factor for premature vascular disease and thrombosis, induces tissue factor activity in endothelial cells. Arterioscler Thromb 1993; 13: 1327-33.
    CrossrefPubMedGoogle Scholar
  • 8 Diquelou A, Dupouy D, Gaspin D, Constans J, Sie P, Boneu B, Sakariassen KS, Cadroy Y. Relationship between endothelial tissue factor and thrombogenesis under blood flow conditions. Thromb Haemost 1995; 74: 778-83.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 9 Berliner J, Leitinger N, Watson A, Huber J, Fogelman A, Navab M. Oxidized lipids in atherogenesis: formation, destruction and action. Thromb Haemost 1997; 78: 195-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 10 Quinn MT, Parthasarathy S, Fong GL, Steinberg D. Oxidatively modified low density lipoproteins: a potential role in recruitment and retention of monocyte/macrophages during atherogenesis. Proc Natl Acad Sci USA 1987; 84: 2995-8.
    CrossrefPubMedGoogle Scholar
  • 11 Frostegard J, Nilsson J, Haegerstrand A, Hamsten A, Wigzell H, Gidlund M. Oxidized low density lipoprotein induces differentiation and adhesion of human monocytes and the monocytic cell line U 937. Proc Natl Acad Sci USA 1990; 87: 904-8.
    CrossrefPubMedGoogle Scholar
  • 12 Weber C, Erl W, Weber PC. Enhancement of monocyte adhesion to endothelial cells by oxidatively modified low-density lipoprotein is mediated by activation of CD11b. Biochem Biophys Res Commol/Lun 1995; 206: 621-8.
    PubMedGoogle Scholar
  • 13 Erl W, Weber PC, Weber C. Monocytic cell adhesion to endothelial cells stimulated by oxidized low density lipoprotein is mediated by distinct endothelial ligands. Atherosclerosis 1998; 136: 297-303.
    CrossrefPubMedGoogle Scholar
  • 14 Bombeli T, Schwartz BR, Harlan JM. Adhesion of activated platelets to endothelial cells: evidence for a GPIIbIIIa-dependent bridging mechanism and novel roles for endothelial intercellular adhesion molecule 1 (ICAM-1), αvβ3 integrin and GPIba. J Exp Med 1998; 187: 329-39.
    CrossrefPubMedGoogle Scholar
  • 15 Gawaz M, Neumann FJ, Dickfeld T, Reininger A, Adelsberger H, Gebhardt A, Schomig A. Vitronectin receptor (αvβ3) mediates platelet adhesion to the luminal aspect of endothelial cells. Implications for reperfusion in acute myocardial infarction. Circulation 1997; 96: 1809-17.
    CrossrefPubMedGoogle Scholar
  • 16 Havel RJ, Eder HA, Bragdon JH. Distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum. J Clin Invest 1955; 34: 1345-53.
    CrossrefPubMedGoogle Scholar
  • 17 Buege JA, Aust SD. Microsomal lipid peroxidation. Methods Enzymol 1976; 52: 302-10.
    PubMedGoogle Scholar
  • 18 Varon D, Dardik R, Shenkamn B, Kotev-Emeth S, Farzame N, Tamarin I, Savion N. A new method for quantitative analysis of whole blood platelet interaction with extracellular matrix under flow conditions. Thromb Res 1997; 85: 283-94.
    CrossrefPubMedGoogle Scholar
  • 19 Varon D, Lashevski I, Brenner B, Beyar R, Lanir N, Tamarin I, Savion N. Cone and plate(let) analyzer: Monitoring glycoprotein IIb/IIIa antagonists and von Willebrand disease replacement therapy by testing platelet deposition under flow conditions. Amer Heart J 1998; 135: S187-93.
    CrossrefPubMedGoogle Scholar
  • 20 Lavee J, Martinowitz U, Mohr R, Goor D, Golan M, Langsam J, Malik Z, Savion N. The effect of transfusion of fresh whole blood versus platelet concentrates after cardiac operations. J Thorac Cardiovasc Surg 1989; 97: 204-12.
    PubMedGoogle Scholar
  • 21 Weiss HJ, Tschopp TB, Rogers J, Brand H. Studies of platelet 5-hydroxytryptamine (serotonin) in storage pool disease and albinism. J Clin Invest 1974; 54: 421-32.
    CrossrefPubMedGoogle Scholar
  • 22 Kokame K, Kato H, Miyata T. Homocysteine – respondent genes in vascular endothelial cells identified by differential display analysis. J Biol Chem 1996; 271: 29659-65.
    CrossrefPubMedGoogle Scholar
  • 23 Dardik R, Savion N, Kaufmann Y, Varon D. Thrombin promotes plateletmediated melanoma cell adhesion to endothelial cells under flow conditions: role of platelet glycoproteins P-selectin and GPIIb-IIIa. Br J Cancer 1998; 77: 2069-75.
    CrossrefPubMedGoogle Scholar
  • 24 Kaplan JE, Moon DG, Weston LK, Minnear FL, Del Vecchio PJ, Shepard JM, Fenton II JW. Platelets adhere to thrombin-treated endothelial cells in vitro. Am J Physiol 1989; 257: H423-33.
    PubMedGoogle Scholar
  • 25 Li JL, Podolsky RS, Rohrer MJ, Cutler BS, Massie MT, Barnard MR, Michenson MD. Adhesion of activated platelets to venous endothelial cells is mediated via GPIIb/IIIa. J Surg Res 1996; 61: 543-58.
    CrossrefPubMedGoogle Scholar
  • 26 Meyers KM, Holmsen Het, Seachord CL. Comparative study of platelet dense granule constituents. Am J Physiol 1982; 243: 454.
    PubMedGoogle Scholar
  • 27 Weis JR, Pitas RE, Wilson BD, Rodgers GM. Oxidized LDL increases cultured human endothelial cell tissue factor activity and reduces protein C activation. Faseb J 1991; 05: 2459-65.
    CrossrefPubMedGoogle Scholar
  • 28 Tam SH, Sassoli PM, Jordan RE, Nakada M. Abciximab (ReoPro, chimeric 7E3 Fab) demonstrates equivalent affinity and functional blockade of glycoprotein IIb/IIIa and αvβ3 integrins. Circulation 1998; 98: 1085-91.
    CrossrefPubMedGoogle Scholar
  • 29 Reininger AJ, Korndorfer MA, Wurzinger LJ. Adhesion of ADP-activated platelets to intact endothelium under stagnation point flow in vitro is mediated by the integrin IIbβ3 . Thromb Haemost 1998; 79: 998-1003.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 De Jong SC, Stehouwer CDA, van den Berg M, Vischer UM, Rauwerda JA, Emeis JJ. Endothelial marker proteins in hyperhomocysteinemia. Thromb Haemost 1997; 78: 1332-7.
    Article in Thieme ConnectPubMedGoogle Scholar