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. 1991 Mar;138(3):601–607.

Minimally oxidized low-density lipoprotein induces tissue factor expression in cultured human endothelial cells.

T A Drake 1, K Hannani 1, H H Fei 1, S Lavi 1, J A Berliner 1
PMCID: PMC1886294  PMID: 2000938

Abstract

Oxidatively modified low-density lipoprotein is present in atherosclerotic lesions and has been proposed to play an important role in atherogenesis through its biologic effects on vascular cells. This study examined the effects of minimally oxidized preparations of LDL (MM-LDL) on tissue factor (TF) expression by cultured human endothelial cells. Low-density lipoprotein purified from normal donors was modified by exposure to iron or by prolonged storage, resulting in levels of thiobarbituric acid-reacting substances of approximately 2.5 to 4 nmoles/mg cholesterol. Preparations had less than 2.5 pg of endotoxin per microgram LDL and had no intrinsic procoagulant activity. This form of modified but not native LDL induced TF expression in endothelial cells in a time- and dose-dependent manner. Peak TF coagulant activity in cells exposed to 40 micrograms/ml MM-LDL were observed at 4 to 6 hours, and ranged from 50 to 500 pg/10(5) cells, compared with less than 10 pg/10(5) cells exposed to native LDL. Northern blot analysis showed TF mRNA levels to increase approximately 30-fold with exposure to MM-LDL for 2 hours. Induction of TF activity was dependent on the concentration of MM-LDL from 1 microgram/ml to 80 micrograms/ml, a range in which cell viability and morphology were unaffected. The findings suggest that minimally oxidized LDL may be a local mediator promoting thrombosis in atherosclerotic lesions.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bar-Shavit R., Wilner G. D. Mediation of cellular events by thrombin. Int Rev Exp Pathol. 1986;29:213–241. [PubMed] [Google Scholar]
  2. Berliner J. A., Territo M. C., Sevanian A., Ramin S., Kim J. A., Bamshad B., Esterson M., Fogelman A. M. Minimally modified low density lipoprotein stimulates monocyte endothelial interactions. J Clin Invest. 1990 Apr;85(4):1260–1266. doi: 10.1172/JCI114562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyd H. C., Gown A. M., Wolfbauer G., Chait A. Direct evidence for a protein recognized by a monoclonal antibody against oxidatively modified LDL in atherosclerotic lesions from a Watanabe heritable hyperlipidemic rabbit. Am J Pathol. 1989 Nov;135(5):815–825. [PMC free article] [PubMed] [Google Scholar]
  4. Carew T. E., Schwenke D. C., Steinberg D. Antiatherogenic effect of probucol unrelated to its hypocholesterolemic effect: evidence that antioxidants in vivo can selectively inhibit low density lipoprotein degradation in macrophage-rich fatty streaks and slow the progression of atherosclerosis in the Watanabe heritable hyperlipidemic rabbit. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7725–7729. doi: 10.1073/pnas.84.21.7725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Conway E. M., Bach R., Rosenberg R. D., Konigsberg W. H. Tumor necrosis factor enhances expression of tissue factor mRNA in endothelial cells. Thromb Res. 1989 Feb 1;53(3):231–241. doi: 10.1016/0049-3848(89)90098-4. [DOI] [PubMed] [Google Scholar]
  7. Crossman D. C., Carr D. P., Tuddenham E. G., Pearson J. D., McVey J. H. The regulation of tissue factor mRNA in human endothelial cells in response to endotoxin or phorbol ester. J Biol Chem. 1990 Jun 15;265(17):9782–9787. [PubMed] [Google Scholar]
  8. Drake T. A., Morrissey J. H., Edgington T. S. Selective cellular expression of tissue factor in human tissues. Implications for disorders of hemostasis and thrombosis. Am J Pathol. 1989 May;134(5):1087–1097. [PMC free article] [PubMed] [Google Scholar]
  9. Drake T. A., Pang M. Effects of interleukin-1, lipopolysaccharide, and streptococci on procoagulant activity of cultured human cardiac valve endothelial and stromal cells. Infect Immun. 1989 Feb;57(2):507–512. doi: 10.1128/iai.57.2.507-512.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  11. Fox P. L., Chisolm G. M., DiCorleto P. E. Lipoprotein-mediated inhibition of endothelial cell production of platelet-derived growth factor-like protein depends on free radical lipid peroxidation. J Biol Chem. 1987 May 5;262(13):6046–6054. [PubMed] [Google Scholar]
  12. Frostegård 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 U937. Proc Natl Acad Sci U S A. 1990 Feb;87(3):904–908. doi: 10.1073/pnas.87.3.904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Haberland M. E., Fong D., Cheng L. Malondialdehyde-altered protein occurs in atheroma of Watanabe heritable hyperlipidemic rabbits. Science. 1988 Jul 8;241(4862):215–218. doi: 10.1126/science.2455346. [DOI] [PubMed] [Google Scholar]
  14. Harpold M. M., Evans R. M., Salditt-Georgieff M., Darnell J. E. Production of mRNA in Chinese hamster cells: relationship of the rate of synthesis to the cytoplasmic concentration of nine specific mRNA sequences. Cell. 1979 Aug;17(4):1025–1035. doi: 10.1016/0092-8674(79)90341-6. [DOI] [PubMed] [Google Scholar]
  15. Hessler J. R., Morel D. W., Lewis L. J., Chisolm G. M. Lipoprotein oxidation and lipoprotein-induced cytotoxicity. Arteriosclerosis. 1983 May-Jun;3(3):215–222. doi: 10.1161/01.atv.3.3.215. [DOI] [PubMed] [Google Scholar]
  16. Jürgens G., Hoff H. F., Chisolm G. M., 3rd, Esterbauer H. Modification of human serum low density lipoprotein by oxidation--characterization and pathophysiological implications. Chem Phys Lipids. 1987 Nov-Dec;45(2-4):315–336. doi: 10.1016/0009-3084(87)90070-3. [DOI] [PubMed] [Google Scholar]
  17. Kosugi K., Morel D. W., DiCorleto P. E., Chisolm G. M. Toxicity of oxidized low-density lipoprotein to cultured fibroblasts is selective for S phase of the cell cycle. J Cell Physiol. 1987 Mar;130(3):311–320. doi: 10.1002/jcp.1041300302. [DOI] [PubMed] [Google Scholar]
  18. Lee D. M. Malondialdehyde formation in stored plasma. Biochem Biophys Res Commun. 1980 Aug 29;95(4):1663–1672. doi: 10.1016/s0006-291x(80)80090-8. [DOI] [PubMed] [Google Scholar]
  19. Levy G. A., Schwartz B. S., Curtiss L. K., Edgington T. S. Plasma lipoprotein induction and suppression of the generation of cellular procoagulant activity in vitro. J Clin Invest. 1981 Jun;67(6):1614–1622. doi: 10.1172/JCI110196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Morrissey J. H., Fair D. S., Edgington T. S. Monoclonal antibody analysis of purified and cell-associated tissue factor. Thromb Res. 1988 Nov 1;52(3):247–261. doi: 10.1016/0049-3848(88)90084-9. [DOI] [PubMed] [Google Scholar]
  21. Morrissey J. H., Fakhrai H., Edgington T. S. Molecular cloning of the cDNA for tissue factor, the cellular receptor for the initiation of the coagulation protease cascade. Cell. 1987 Jul 3;50(1):129–135. doi: 10.1016/0092-8674(87)90669-6. [DOI] [PubMed] [Google Scholar]
  22. Morrissey J. H., Revak D., Tejada P., Fair D. S., Edgington T. S. Resolution of monomeric and heterodimeric forms of tissue factor, the high-affinity cellular receptor for factor VII. Thromb Res. 1988 May 15;50(4):481–493. doi: 10.1016/0049-3848(88)90197-1. [DOI] [PubMed] [Google Scholar]
  23. Munro J. M., Cotran R. S. The pathogenesis of atherosclerosis: atherogenesis and inflammation. Lab Invest. 1988 Mar;58(3):249–261. [PubMed] [Google Scholar]
  24. Nemerson Y. Tissue factor and hemostasis. Blood. 1988 Jan;71(1):1–8. [PubMed] [Google Scholar]
  25. Pober J. S. Warner-Lambert/Parke-Davis award lecture. Cytokine-mediated activation of vascular endothelium. Physiology and pathology. Am J Pathol. 1988 Dec;133(3):426–433. [PMC free article] [PubMed] [Google Scholar]
  26. Rajavashisth T. B., Andalibi A., Territo M. C., Berliner J. A., Navab M., Fogelman A. M., Lusis A. J. Induction of endothelial cell expression of granulocyte and macrophage colony-stimulating factors by modified low-density lipoproteins. Nature. 1990 Mar 15;344(6263):254–257. doi: 10.1038/344254a0. [DOI] [PubMed] [Google Scholar]
  27. Rajavashisth T. B., Eng R., Shadduck R. K., Waheed A., Ben-Avram C. M., Shively J. E., Lusis A. J. Cloning and tissue-specific expression of mouse macrophage colony-stimulating factor mRNA. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1157–1161. doi: 10.1073/pnas.84.5.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rodgers G. M. Hemostatic properties of normal and perturbed vascular cells. FASEB J. 1988 Feb;2(2):116–123. doi: 10.1096/fasebj.2.2.3277885. [DOI] [PubMed] [Google Scholar]
  29. Scarpati E. M., Sadler J. E. Regulation of endothelial cell coagulant properties. Modulation of tissue factor, plasminogen activator inhibitors, and thrombomodulin by phorbol 12-myristate 13-acetate and tumor necrosis factor. J Biol Chem. 1989 Dec 5;264(34):20705–20713. [PubMed] [Google Scholar]
  30. Schuff-Werner P., Claus G., Armstrong V. W., Köstering H., Seidel D. Enhanced procoagulatory activity (PCA) of human monocytes/macrophages after in vitro stimulation with chemically modified LDL. Atherosclerosis. 1989 Aug;78(2-3):109–112. doi: 10.1016/0021-9150(89)90214-1. [DOI] [PubMed] [Google Scholar]
  31. Schwartz B. S., Levy G. A., Curtiss L. K., Fair D. S., Edgington T. S. Plasma lipoprotein induction and suppression of the generation of cellular procoagulant activity in vitro: two procoagulant activities are produced by peripheral blood mononuclear cells. J Clin Invest. 1981 Jun;67(6):1650–1658. doi: 10.1172/JCI110201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wilcox J. N., Smith K. M., Schwartz S. M., Gordon D. Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2839–2843. doi: 10.1073/pnas.86.8.2839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ylä-Herttuala S., Palinski W., Rosenfeld M. E., Parthasarathy S., Carew T. E., Butler S., Witztum J. L., Steinberg D. Evidence for the presence of oxidatively modified low density lipoprotein in atherosclerotic lesions of rabbit and man. J Clin Invest. 1989 Oct;84(4):1086–1095. doi: 10.1172/JCI114271. [DOI] [PMC free article] [PubMed] [Google Scholar]

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