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. 1988 Apr;85(7):2344–2348. doi: 10.1073/pnas.85.7.2344

Cholesteryl ester accumulation in macrophages incubated with low density lipoprotein pretreated with cigarette smoke extract.

M Yokode 1, T Kita 1, H Arai 1, C Kawai 1, S Narumiya 1, M Fujiwara 1
PMCID: PMC279988  PMID: 3353382

Abstract

Although cigarette smoking is one of the major risk factors for atherosclerosis and coronary heart disease, the precise mechanisms of its adverse effects have not been fully elucidated. We incubated low density lipoprotein (LDL) with cigarette smoke (CS) extract and examined the incorporation of the lipoprotein by macrophages in vitro. When incubated with macrophages, LDL pretreated with CS extract (100 micrograms/ml) stimulated cholesteryl [14C]oleate synthesis approximately equal to 12.5-fold that with unmodified LDL and transformed macrophages to cells rich in lipid droplets positively stained with oil red O. Enhancement in cholesteryl ester synthesis was dependent on the concentration of CS-modified LDL and exhibited saturation kinetics. When subjected to electrophoreses, CS-modified LDL migrated to a more anionic position than did unmodified LDL and showed extensive fragmentation of apolipoprotein B. This LDL modification depended upon the incubation time and concentration of the CS extract. Superoxide dismutase inhibited modification of LDL by 52%, suggesting that superoxide anion is, at least in part, involved. These results suggest that CS extract alters LDL into a form recognized and incorporated by macrophages. Such modification if it occurs in vivo, could explain the increased incidence of atherosclerosis and coronary heart disease in smokers.

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Selected References

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

  1. Ball K., Turner R. Smoking and the heart. The basis for action. Lancet. 1974 Oct 5;2(7884):822–826. doi: 10.1016/s0140-6736(74)91083-6. [DOI] [PubMed] [Google Scholar]
  2. Basu S. K., Anderson R. G., Goldstein J. L., Brown M. S. Metabolism of cationized lipoproteins by human fibroblasts. Biochemical and morphologic correlations. J Cell Biol. 1977 Jul;74(1):119–135. doi: 10.1083/jcb.74.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Basu S. K., Goldstein J. L., Anderson G. W., Brown M. S. Degradation of cationized low density lipoprotein and regulation of cholesterol metabolism in homozygous familial hypercholesterolemia fibroblasts. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3178–3182. doi: 10.1073/pnas.73.9.3178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brischetto C. S., Connor W. E., Connor S. L., Matarazzo J. D. Plasma lipid and lipoprotein profiles of cigarette smokers from randomly selected families: enhancement of hyperlipidemia and depression of high-density lipoprotein. Am J Cardiol. 1983 Oct 1;52(7):675–680. doi: 10.1016/0002-9149(83)90396-x. [DOI] [PubMed] [Google Scholar]
  5. Brown M. S., Basu S. K., Falck J. R., Ho Y. K., Goldstein J. L. The scavenger cell pathway for lipoprotein degradation: specificity of the binding site that mediates the uptake of negatively-charged LDL by macrophages. J Supramol Struct. 1980;13(1):67–81. doi: 10.1002/jss.400130107. [DOI] [PubMed] [Google Scholar]
  6. Brown M. S., Goldstein J. L., Krieger M., Ho Y. K., Anderson R. G. Reversible accumulation of cholesteryl esters in macrophages incubated with acetylated lipoproteins. J Cell Biol. 1979 Sep;82(3):597–613. doi: 10.1083/jcb.82.3.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brown M. S., Goldstein J. L. Lipoprotein metabolism in the macrophage: implications for cholesterol deposition in atherosclerosis. Annu Rev Biochem. 1983;52:223–261. doi: 10.1146/annurev.bi.52.070183.001255. [DOI] [PubMed] [Google Scholar]
  8. Buja L. M., Kita T., Goldstein J. L., Watanabe Y., Brown M. S. Cellular pathology of progressive atherosclerosis in the WHHL rabbit. An animal model of familial hypercholesterolemia. Arteriosclerosis. 1983 Jan-Feb;3(1):87–101. doi: 10.1161/01.atv.3.1.87. [DOI] [PubMed] [Google Scholar]
  9. Cryer P. E., Haymond M. W., Santiago J. V., Shah S. D. Norepinephrine and epinephrine release and adrenergic mediation of smoking-associated hemodynamic and metabolic events. N Engl J Med. 1976 Sep 9;295(11):573–577. doi: 10.1056/NEJM197609092951101. [DOI] [PubMed] [Google Scholar]
  10. Fogelman A. M., Shechter I., Seager J., Hokom M., Child J. S., Edwards P. A. Malondialdehyde alteration of low density lipoproteins leads to cholesteryl ester accumulation in human monocyte-macrophages. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2214–2218. doi: 10.1073/pnas.77.4.2214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gerrity R. G. The role of the monocyte in atherogenesis: II. Migration of foam cells from atherosclerotic lesions. Am J Pathol. 1981 May;103(2):191–200. [PMC free article] [PubMed] [Google Scholar]
  12. Goldstein J. L., Ho Y. K., Basu S. K., Brown M. S. Binding site on macrophages that mediates uptake and degradation of acetylated low density lipoprotein, producing massive cholesterol deposition. Proc Natl Acad Sci U S A. 1979 Jan;76(1):333–337. doi: 10.1073/pnas.76.1.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gutteridge J. M. The role of superoxide and hydroxyl radicals in phospholipid peroxidation catalysed by iron salts. FEBS Lett. 1982 Dec 27;150(2):454–458. doi: 10.1016/0014-5793(82)80788-6. [DOI] [PubMed] [Google Scholar]
  14. HAVEL R. J., EDER H. A., BRAGDON J. H. The distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum. J Clin Invest. 1955 Sep;34(9):1345–1353. doi: 10.1172/JCI103182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Heinecke J. W., Baker L., Rosen H., Chait A. Superoxide-mediated modification of low density lipoprotein by arterial smooth muscle cells. J Clin Invest. 1986 Mar;77(3):757–761. doi: 10.1172/JCI112371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Heinecke J. W., Rosen H., Chait A. Iron and copper promote modification of low density lipoprotein by human arterial smooth muscle cells in culture. J Clin Invest. 1984 Nov;74(5):1890–1894. doi: 10.1172/JCI111609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kita T., Nagano Y., Yokode M., Ishii K., Kume N., Ooshima A., Yoshida H., Kawai C. Probucol prevents the progression of atherosclerosis in Watanabe heritable hyperlipidemic rabbit, an animal model for familial hypercholesterolemia. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5928–5931. doi: 10.1073/pnas.84.16.5928. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kita T., Yokode M., Watanabe Y., Narumiya S., Kawai C. Stimulation of cholesteryl ester synthesis in mouse peritoneal macrophages by cholesterol-rich very low density lipoproteins from the Watanabe heritable hyperlipidemic rabbit, an animal model of familial hypercholesterolemia. J Clin Invest. 1986 May;77(5):1460–1465. doi: 10.1172/JCI112458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. LANGE R. Inhibiting effect of tobacco smoke on some crystalline enzymes. Science. 1961 Jul 7;134(3471):52–53. doi: 10.1126/science.134.3471.52. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Nakayama T., Kaneko M., Kodama M., Nagata C. Cigarette smoke induces DNA single-strand breaks in human cells. Nature. 1985 Apr 4;314(6010):462–464. doi: 10.1038/314462a0. [DOI] [PubMed] [Google Scholar]
  23. Nakayama T., Kodama M., Nagata C. Generation of hydrogen peroxide and superoxide anion radical from cigarette smoke. Gan. 1984 Feb;75(2):95–98. [PubMed] [Google Scholar]
  24. Noble R. P. Electrophoretic separation of plasma lipoproteins in agarose gel. J Lipid Res. 1968 Nov;9(6):693–700. [PubMed] [Google Scholar]
  25. Parthasarathy S., Steinbrecher U. P., Barnett J., Witztum J. L., Steinberg D. Essential role of phospholipase A2 activity in endothelial cell-induced modification of low density lipoprotein. Proc Natl Acad Sci U S A. 1985 May;82(9):3000–3004. doi: 10.1073/pnas.82.9.3000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Parthasarathy S., Young S. G., Witztum J. L., Pittman R. C., Steinberg D. Probucol inhibits oxidative modification of low density lipoprotein. J Clin Invest. 1986 Feb;77(2):641–644. doi: 10.1172/JCI112349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schuh J., Fairclough G. F., Jr, Haschemeyer R. H. Oxygen-mediated heterogeneity of apo-low-density lipoprotein. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3173–3177. doi: 10.1073/pnas.75.7.3173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Seltzer C. C. Smoking and coronary heart disease: what are we to believe? Am Heart J. 1980 Sep;100(3):275–280. doi: 10.1016/0002-8703(80)90139-8. [DOI] [PubMed] [Google Scholar]
  29. Steinbrecher U. P. Oxidation of human low density lipoprotein results in derivatization of lysine residues of apolipoprotein B by lipid peroxide decomposition products. J Biol Chem. 1987 Mar 15;262(8):3603–3608. [PubMed] [Google Scholar]
  30. Steinbrecher U. P., Parthasarathy S., Leake D. S., Witztum J. L., Steinberg D. Modification of low density lipoprotein by endothelial cells involves lipid peroxidation and degradation of low density lipoprotein phospholipids. Proc Natl Acad Sci U S A. 1984 Jun;81(12):3883–3887. doi: 10.1073/pnas.81.12.3883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wald N., Howard S., Smith P. G., Kjeldsen K. Association between atherosclerotic diseases and carboxyhaemoglobin levels in tobacco smokers. Br Med J. 1973 Mar 31;1(5856):761–765. doi: 10.1136/bmj.1.5856.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Winniford M. D., Wheelan K. R., Kremers M. S., Ugolini V., van den Berg E., Jr, Niggemann E. H., Jansen D. E., Hillis L. D. Smoking-induced coronary vasoconstriction in patients with atherosclerotic coronary artery disease: evidence for adrenergically mediated alterations in coronary artery tone. Circulation. 1986 Apr;73(4):662–667. doi: 10.1161/01.cir.73.4.662. [DOI] [PubMed] [Google Scholar]
  33. Yagi K. A simple fluorometric assay for lipoperoxide in blood plasma. Biochem Med. 1976 Apr;15(2):212–216. doi: 10.1016/0006-2944(76)90049-1. [DOI] [PubMed] [Google Scholar]
  34. Yokode M., Kita T., Narumiya S., Kawai C. Arachidonate metabolism during foam cell transformation of macrophages. Adv Prostaglandin Thromboxane Leukot Res. 1987;17A:229–232. [PubMed] [Google Scholar]

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