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. 1991 Feb;87(2):597–601. doi: 10.1172/JCI115035

Preservation of the endogenous antioxidants in low density lipoprotein by ascorbate but not probucol during oxidative modification.

I Jialal 1, S M Grundy 1
PMCID: PMC296348  PMID: 1991843

Abstract

Several lines of evidence indicate that the oxidative modification of low density lipoproteins (LDL) may provide an important link between plasma LDL and the genesis of the atherosclerotic lesion. Ascorbate is an important water-soluble, chain-breaking antioxidant in humans. Probucol, a lipid-soluble antioxidant drug has been shown to retard the progression of atherosclerosis. The aim of the present study was to compare the effects of probucol and physiologic levels of ascorbate on the oxidative modification of LDL in both a cell-free (2.5 microM Cu++ in phosphate-buffered saline) and cellular system (human monocyte macrophages in Ham's F-10 medium). Both ascorbate and probucol inhibited the oxidative modification of LDL in both systems to a similar degree as evidenced by the thiobarbituric acid-reacting substance activity, electrophoretic mobility, and degradation by macrophages. However, whereas co-incubation with physiologic levels of ascorbate resulted in a substantial preservation of the alpha-tocopherol, gamma-tocopherol, and beta-carotene of the LDL, probucol in concentrations ranging from 10 to 80 microM failed to protect these antioxidants. Thus, in addition to being as potent as probucol in inhibiting the oxidation of LDL, ascorbate in contrast preserves the endogenous antioxidants in the LDL.

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Selected References

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  1. Aviram M., Bierman E. L., Chait A. Modification of low density lipoprotein by lipoprotein lipase or hepatic lipase induces enhanced uptake and cholesterol accumulation in cells. J Biol Chem. 1988 Oct 25;263(30):15416–15422. [PubMed] [Google Scholar]
  2. Bendich A., D'Apolito P., Gabriel E., Machlin L. J. Interaction of dietary vitamin C and vitamin E on guinea pig immune responses to mitogens. J Nutr. 1984 Sep;114(9):1588–1593. doi: 10.1093/jn/114.9.1588. [DOI] [PubMed] [Google Scholar]
  3. Bierman E. L., Stein O., Stein Y. Lipoprotein uptake and metabolism by rat aortic smooth muscle cells in tissue culture. Circ Res. 1974 Jul;35(1):136–150. doi: 10.1161/01.res.35.1.136. [DOI] [PubMed] [Google Scholar]
  4. Bodannes R. S., Chan P. C. Ascorbic acid as a scavenger of singlet oxygen. FEBS Lett. 1979 Sep 15;105(2):195–196. doi: 10.1016/0014-5793(79)80609-2. [DOI] [PubMed] [Google Scholar]
  5. Buckley M. M., Goa K. L., Price A. H., Brogden R. N. Probucol. A reappraisal of its pharmacological properties and therapeutic use in hypercholesterolaemia. Drugs. 1989 Jun;37(6):761–800. doi: 10.2165/00003495-198937060-00002. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Dachet C., Jacotot B., Buxtorf J. C. The hypolipidemic action of probucol. Drug transport and lipoprotein composition in type IIa hyperlipoproteinemia. Atherosclerosis. 1985 Dec;58(1-3):261–268. doi: 10.1016/0021-9150(85)90071-1. [DOI] [PubMed] [Google Scholar]
  8. Doba T., Burton G. W., Ingold K. U. Antioxidant and co-antioxidant activity of vitamin C. The effect of vitamin C, either alone or in the presence of vitamin E or a water-soluble vitamin E analogue, upon the peroxidation of aqueous multilamellar phospholipid liposomes. Biochim Biophys Acta. 1985 Jul 9;835(2):298–303. doi: 10.1016/0005-2760(85)90285-1. [DOI] [PubMed] [Google Scholar]
  9. Dubick M. A., Hunter G. C., Casey S. M., Keen C. L. Aortic ascorbic acid, trace elements, and superoxide dismutase activity in human aneurysmal and occlusive disease. Proc Soc Exp Biol Med. 1987 Feb;184(2):138–143. doi: 10.3181/00379727-184-42457. [DOI] [PubMed] [Google Scholar]
  10. Esterbauer H., Striegl G., Puhl H., Rotheneder M. Continuous monitoring of in vitro oxidation of human low density lipoprotein. Free Radic Res Commun. 1989;6(1):67–75. doi: 10.3109/10715768909073429. [DOI] [PubMed] [Google Scholar]
  11. Fellin R., Gasparotto A., Valerio G., Baiocchi M. R., Padrini R., Lamon S., Vitale E., Baggio G., Crepaldi G. Effect of probucol treatment on lipoprotein cholesterol and drug levels in blood and lipoproteins in familial hypercholesterolemia. Atherosclerosis. 1986 Jan;59(1):47–56. doi: 10.1016/0021-9150(86)90032-8. [DOI] [PubMed] [Google Scholar]
  12. Frei B., England L., Ames B. N. Ascorbate is an outstanding antioxidant in human blood plasma. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6377–6381. doi: 10.1073/pnas.86.16.6377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gey K. F., Brubacher G. B., Stähelin H. B. Plasma levels of antioxidant vitamins in relation to ischemic heart disease and cancer. Am J Clin Nutr. 1987 May;45(5 Suppl):1368–1377. doi: 10.1093/ajcn/45.5.1368. [DOI] [PubMed] [Google Scholar]
  14. Grundy S. M., Vega G. L. Influence of mevinolin on metabolism of low density lipoproteins in primary moderate hypercholesterolemia. J Lipid Res. 1985 Dec;26(12):1464–1475. [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Henriksen T., Mahoney E. M., Steinberg D. Enhanced macrophage degradation of biologically modified low density lipoprotein. Arteriosclerosis. 1983 Mar-Apr;3(2):149–159. doi: 10.1161/01.atv.3.2.149. [DOI] [PubMed] [Google Scholar]
  18. Henriksen T., Mahoney E. M., Steinberg D. Enhanced macrophage degradation of low density lipoprotein previously incubated with cultured endothelial cells: recognition by receptors for acetylated low density lipoproteins. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6499–6503. doi: 10.1073/pnas.78.10.6499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hiramatsu K., Rosen H., Heinecke J. W., Wolfbauer G., Chait A. Superoxide initiates oxidation of low density lipoprotein by human monocytes. Arteriosclerosis. 1987 Jan-Feb;7(1):55–60. doi: 10.1161/01.atv.7.1.55. [DOI] [PubMed] [Google Scholar]
  20. Jialal I., Vega G. L., Grundy S. M. Physiologic levels of ascorbate inhibit the oxidative modification of low density lipoprotein. Atherosclerosis. 1990 Jun;82(3):185–191. doi: 10.1016/0021-9150(90)90039-l. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. 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]
  23. Ku G., Schroeder K., Schmidt L. F., Jackson R. L., Doherty N. S. Probucol does not alter acetylated low density lipoprotein uptake by murine peritoneal macrophages. Atherosclerosis. 1990 Jan;80(3):191–197. doi: 10.1016/0021-9150(90)90026-f. [DOI] [PubMed] [Google Scholar]
  24. Levine M. New concepts in the biology and biochemistry of ascorbic acid. N Engl J Med. 1986 Apr 3;314(14):892–902. doi: 10.1056/NEJM198604033141407. [DOI] [PubMed] [Google Scholar]
  25. McLean L. R., Hagaman K. A. Effect of probucol on the physical properties of low-density lipoproteins oxidized by copper. Biochemistry. 1989 Jan 10;28(1):321–327. doi: 10.1021/bi00427a043. [DOI] [PubMed] [Google Scholar]
  26. Morel D. W., Hessler J. R., Chisolm G. M. Low density lipoprotein cytotoxicity induced by free radical peroxidation of lipid. J Lipid Res. 1983 Aug;24(8):1070–1076. [PubMed] [Google Scholar]
  27. Nagano Y., Kita T., Yokode M., Ishii K., Kume N., Otani H., Arai H., Kawai C. Probucol does not affect lipoprotein metabolism in macrophages of Watanabe heritable hyperlipidemic rabbits. Arteriosclerosis. 1989 Jul-Aug;9(4):453–461. doi: 10.1161/01.atv.9.4.453. [DOI] [PubMed] [Google Scholar]
  28. Niki E. Antioxidants in relation to lipid peroxidation. Chem Phys Lipids. 1987 Jul-Sep;44(2-4):227–253. doi: 10.1016/0009-3084(87)90052-1. [DOI] [PubMed] [Google Scholar]
  29. Nishikimi M. Oxidation of ascorbic acid with superoxide anion generated by the xanthine-xanthine oxidase system. Biochem Biophys Res Commun. 1975 Mar 17;63(2):463–468. doi: 10.1016/0006-291x(75)90710-x. [DOI] [PubMed] [Google Scholar]
  30. Noble R. P. Electrophoretic separation of plasma lipoproteins in agarose gel. J Lipid Res. 1968 Nov;9(6):693–700. [PubMed] [Google Scholar]
  31. Packer J. E., Slater T. F., Willson R. L. Direct observation of a free radical interaction between vitamin E and vitamin C. Nature. 1979 Apr 19;278(5706):737–738. doi: 10.1038/278737a0. [DOI] [PubMed] [Google Scholar]
  32. Palinski W., Rosenfeld M. E., Ylä-Herttuala S., Gurtner G. C., Socher S. S., Butler S. W., Parthasarathy S., Carew T. E., Steinberg D., Witztum J. L. Low density lipoprotein undergoes oxidative modification in vivo. Proc Natl Acad Sci U S A. 1989 Feb;86(4):1372–1376. doi: 10.1073/pnas.86.4.1372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Parthasarathy S., Printz D. J., Boyd D., Joy L., Steinberg D. Macrophage oxidation of low density lipoprotein generates a modified form recognized by the scavenger receptor. Arteriosclerosis. 1986 Sep-Oct;6(5):505–510. doi: 10.1161/01.atv.6.5.505. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Quinn M. T., Parthasarathy S., Fong L. G., Steinberg D. Oxidatively modified low density lipoproteins: a potential role in recruitment and retention of monocyte/macrophages during atherogenesis. Proc Natl Acad Sci U S A. 1987 May;84(9):2995–2998. doi: 10.1073/pnas.84.9.2995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Ramirez J., Flowers N. C. Leukocyte ascorbic acid and its relationship to coronary artery disease in man. Am J Clin Nutr. 1980 Oct;33(10):2079–2087. doi: 10.1093/ajcn/33.10.2079. [DOI] [PubMed] [Google Scholar]
  37. Sauberlich H. E. Human requirements and needs. Vitamin C status: methods and findings. Ann N Y Acad Sci. 1975 Sep 30;258:438–450. doi: 10.1111/j.1749-6632.1975.tb29302.x. [DOI] [PubMed] [Google Scholar]
  38. Steinberg D., Parthasarathy S., Carew T. E., Khoo J. C., Witztum J. L. Beyond cholesterol. Modifications of low-density lipoprotein that increase its atherogenicity. N Engl J Med. 1989 Apr 6;320(14):915–924. doi: 10.1056/NEJM198904063201407. [DOI] [PubMed] [Google Scholar]
  39. Steinbrecher U. P., Witztum J. L., Parthasarathy S., Steinberg D. Decrease in reactive amino groups during oxidation or endothelial cell modification of LDL. Correlation with changes in receptor-mediated catabolism. Arteriosclerosis. 1987 Mar-Apr;7(2):135–143. doi: 10.1161/01.atv.7.2.135. [DOI] [PubMed] [Google Scholar]
  40. Yam L. T., Li C. Y., Crosby W. H. Cytochemical identification of monocytes and granulocytes. Am J Clin Pathol. 1971 Mar;55(3):283–290. doi: 10.1093/ajcp/55.3.283. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. van Hinsbergh V. W., Scheffer M., Havekes L., Kempen H. J. Role of endothelial cells and their products in the modification of low-density lipoproteins. Biochim Biophys Acta. 1986 Aug 14;878(1):49–64. doi: 10.1016/0005-2760(86)90343-7. [DOI] [PubMed] [Google Scholar]

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