Logo of jcinvestThe Journal of Clinical InvestigationCurrent IssueArchiveSubscriptionAbout the Journal
J Clin Invest. 1995 Nov; 96(5): 2322–2330.
PMCID: PMC185883

Oxidatively modified LDL contains phospholipids with platelet-activating factor-like activity and stimulates the growth of smooth muscle cells.


Oxidative modification of lipoproteins is believed to be important in the genesis of atherosclerosis. We established cultures of smooth muscle cells (SMC) and exposed them to native LDL or oxidized LDL. Oxidized LDL, but not native LDL, was mitogenic as measured by incorporation of [3H]-thymidine into DNA. This effect was concentration dependent, averaged 288% of control, and was blocked by a platelet-activating factor (PAF) receptor antagonist. We hypothesized that phospholipids with PAF-like activity were generated during the oxidation of LDL. To test this hypothesis we extracted phospholipids from copper-oxidized LDL and assayed for PAF-like activity. Phospholipids extracted from oxidized LDL and purified by HPLC induced neutrophil adhesion equivalent to PAF (10 nM) and were mitogenic for smooth muscle cells. These effects were not seen with phospholipids extracted from native LDL and were blocked by two structurally different, competitive antagonists of the PAF receptor. The effects of these lipids were also abolished by pretreating them with PAF acetylhydrolase. Finally, we used Chinese hamster ovary cells that had seen stably transfected with a cDNA for the PAF receptor to confirm that phospholipids from oxidized LDL act via this receptor. We found that PAF (control) and the oxidized phospholipids each induced release of arachidonic acid from the transfected cells, but had no effect on wildtype Chinese hamster ovary cells, which lack the PAF receptor. This effect was also blocked by a PAF receptor antagonist. Thus, phospholipids generated during oxidative modification of LDL may participate in atherosclerosis by stimulating SMC proliferation and leukocyte activation.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.2M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Castelli WP. Epidemiology of coronary heart disease: the Framingham study. Am J Med. 1984 Feb 27;76(2A):4–12. [PubMed]
  • Rose G, Shipley M. Plasma cholesterol concentration and death from coronary heart disease: 10 year results of the Whitehall study. Br Med J (Clin Res Ed) 1986 Aug 2;293(6542):306–307. [PMC free article] [PubMed]
  • Stamler J, Wentworth D, Neaton JD. Is relationship between serum cholesterol and risk of premature death from coronary heart disease continuous and graded? Findings in 356,222 primary screenees of the Multiple Risk Factor Intervention Trial (MRFIT). JAMA. 1986 Nov 28;256(20):2823–2828. [PubMed]
  • Brown MS, Goldstein JL. Lipoprotein metabolism in the macrophage: implications for cholesterol deposition in atherosclerosis. Annu Rev Biochem. 1983;52:223–261. [PubMed]
  • Witztum JL, Steinberg D. Role of oxidized low density lipoprotein in atherogenesis. J Clin Invest. 1991 Dec;88(6):1785–1792. [PMC free article] [PubMed]
  • Steinberg D, Parthasarathy S, Carew TE, Khoo JC, Witztum JL. Beyond cholesterol. Modifications of low-density lipoprotein that increase its atherogenicity. N Engl J Med. 1989 Apr 6;320(14):915–924. [PubMed]
  • Henriksen T, Mahoney EM, Steinberg D. Enhanced macrophage degradation of biologically modified low density lipoprotein. Arteriosclerosis. 1983 Mar-Apr;3(2):149–159. [PubMed]
  • Parthasarathy S, Printz DJ, 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. [PubMed]
  • Sparrow CP, Olszewski J. Cellular oxidative modification of low density lipoprotein does not require lipoxygenases. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):128–131. [PMC free article] [PubMed]
  • Björkhem I, Henriksson-Freyschuss A, Breuer O, Diczfalusy U, Berglund L, Henriksson P. The antioxidant butylated hydroxytoluene protects against atherosclerosis. Arterioscler Thromb. 1991 Jan-Feb;11(1):15–22. [PubMed]
  • Sparrow CP, Doebber TW, Olszewski J, Wu MS, Ventre J, Stevens KA, Chao YS. Low density lipoprotein is protected from oxidation and the progression of atherosclerosis is slowed in cholesterol-fed rabbits by the antioxidant N,N'-diphenyl-phenylenediamine. J Clin Invest. 1992 Jun;89(6):1885–1891. [PMC free article] [PubMed]
  • Mao SJ, Yates MT, Parker RA, Chi EM, Jackson RL. Attenuation of atherosclerosis in a modified strain of hypercholesterolemic Watanabe rabbits with use of a probucol analogue (MDL 29,311) that does not lower serum cholesterol. Arterioscler Thromb. 1991 Sep-Oct;11(5):1266–1275. [PubMed]
  • Carew TE, Schwenke DC, 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. [PMC free article] [PubMed]
  • Steinbrecher UP, Lougheed M, Kwan WC, Dirks M. Recognition of oxidized low density lipoprotein by the scavenger receptor of macrophages results from derivatization of apolipoprotein B by products of fatty acid peroxidation. J Biol Chem. 1989 Sep 15;264(26):15216–15223. [PubMed]
  • Goldstein JL, Ho YK, Basu SK, Brown MS. 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. [PMC free article] [PubMed]
  • Stein O, Stein Y. Bovine aortic endothelial cells display macrophage-like properties towards acetylated 125I-labelled low density lipoprotein. Biochim Biophys Acta. 1980 Dec 5;620(3):631–635. [PubMed]
  • Cushing SD, Berliner JA, Valente AJ, Territo MC, Navab M, Parhami F, Gerrity R, Schwartz CJ, Fogelman AM. Minimally modified low density lipoprotein induces monocyte chemotactic protein 1 in human endothelial cells and smooth muscle cells. Proc Natl Acad Sci U S A. 1990 Jul;87(13):5134–5138. [PMC free article] [PubMed]
  • Rajavashisth TB, Andalibi A, Territo MC, Berliner JA, Navab M, Fogelman AM, Lusis AJ. 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. [PubMed]
  • Kugiyama K, Kerns SA, Morrisett JD, Roberts R, Henry PD. Impairment of endothelium-dependent arterial relaxation by lysolecithin in modified low-density lipoproteins. Nature. 1990 Mar 8;344(6262):160–162. [PubMed]
  • Yokoyama M, Hirata K, Miyake R, Akita H, Ishikawa Y, Fukuzaki H. Lysophosphatidylcholine: essential role in the inhibition of endothelium-dependent vasorelaxation by oxidized low density lipoprotein. Biochem Biophys Res Commun. 1990 Apr 16;168(1):301–308. [PubMed]
  • Galle J, Bassenge E, Busse R. Oxidized low density lipoproteins potentiate vasoconstrictions to various agonists by direct interaction with vascular smooth muscle. Circ Res. 1990 May;66(5):1287–1293. [PubMed]
  • Fless GM, Kirchhausen T, Fischer-Dzoga K, Wissler RW, Scanu AM. Serum low density lipoproteins with mitogenic effect on cultured aortic smooth muscle cells. Atherosclerosis. 1982 Feb;41(2-3):171–183. [PubMed]
  • Koschinsky T, Bünting CE, Rütter R, Gries FA. Increased growth stimulation of human vascular cells by serum from patients with primary hyper-LDL-cholesterolemia. Atherosclerosis. 1987 Jan;63(1):7–13. [PubMed]
  • Block LH, Knorr M, Vogt E, Locher R, Vetter W, Groscurth P, Qiao BY, Pometta D, James R, Regenass M, et al. Low density lipoprotein causes general cellular activation with increased phosphatidylinositol turnover and lipoprotein catabolism. Proc Natl Acad Sci U S A. 1988 Feb;85(3):885–889. [PMC free article] [PubMed]
  • Scott-Burden T, Resink TJ, Hahn AW, Baur U, Box RJ, Bühler FR. Induction of growth-related metabolism in human vascular smooth muscle cells by low density lipoprotein. J Biol Chem. 1989 Jul 25;264(21):12582–12589. [PubMed]
  • Sachinidis A, Mengden T, Locher R, Brunner C, Vetter W. Novel cellular activities for low density lipoprotein in vascular smooth muscle cells. Hypertension. 1990 Jun;15(6 Pt 2):704–711. [PubMed]
  • Weisser B, Locher R, Mengden T, Vetter W. Oxidation of low density lipoprotein enhances its potential to increase intracellular free calcium concentration in vascular smooth muscle cells. Arterioscler Thromb. 1992 Feb;12(2):231–236. [PubMed]
  • Resink TJ, Tkachuk VA, Bernhardt J, Bühler FR. Oxidized low density lipoproteins stimulate phosphoinositide turnover in cultured vascular smooth muscle cells. Arterioscler Thromb. 1992 Mar;12(3):278–285. [PubMed]
  • Esterbauer H, Jürgens G, Quehenberger O, Koller E. Autoxidation of human low density lipoprotein: loss of polyunsaturated fatty acids and vitamin E and generation of aldehydes. J Lipid Res. 1987 May;28(5):495–509. [PubMed]
  • Quinn MT, Parthasarathy S, Steinberg D. Lysophosphatidylcholine: a chemotactic factor for human monocytes and its potential role in atherogenesis. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2805–2809. [PMC free article] [PubMed]
  • Zwijsen RM, Japenga SC, Heijen AM, van den Bos RC, Koeman JH. Induction of platelet-derived growth factor chain A gene expression in human smooth muscle cells by oxidized low density lipoproteins. Biochem Biophys Res Commun. 1992 Aug 14;186(3):1410–1416. [PubMed]
  • Locher R, Weisser B, Mengden T, Brunner C, Vetter W. Lysolecithin actions on vascular smooth muscle cells. Biochem Biophys Res Commun. 1992 Feb 28;183(1):156–162. [PubMed]
  • Venable ME, Zimmerman GA, McIntyre TM, Prescott SM. Platelet-activating factor: a phospholipid autacoid with diverse actions. J Lipid Res. 1993 May;34(5):691–702. [PubMed]
  • Stoll LL, Spector AA. Interaction of platelet-activating factor with endothelial and vascular smooth muscle cells in coculture. J Cell Physiol. 1989 May;139(2):253–261. [PubMed]
  • Honda Z, Nakamura M, Miki I, Minami M, Watanabe T, Seyama Y, Okado H, Toh H, Ito K, Miyamoto T, et al. Cloning by functional expression of platelet-activating factor receptor from guinea-pig lung. Nature. 1991 Jan 24;349(6307):342–346. [PubMed]
  • Nakamura M, Honda Z, Izumi T, Sakanaka C, Mutoh H, Minami M, Bito H, Seyama Y, Matsumoto T, Noma M, et al. Molecular cloning and expression of platelet-activating factor receptor from human leukocytes. J Biol Chem. 1991 Oct 25;266(30):20400–20405. [PubMed]
  • Smiley PL, Stremler KE, Prescott SM, Zimmerman GA, McIntyre TM. Oxidatively fragmented phosphatidylcholines activate human neutrophils through the receptor for platelet-activating factor. J Biol Chem. 1991 Jun 15;266(17):11104–11110. [PubMed]
  • Patel KD, Zimmerman GA, Prescott SM, McIntyre TM. Novel leukocyte agonists are released by endothelial cells exposed to peroxide. J Biol Chem. 1992 Jul 25;267(21):15168–15175. [PubMed]
  • Stremler KE, Stafforini DM, Prescott SM, Zimmerman GA, McIntyre TM. An oxidized derivative of phosphatidylcholine is a substrate for the platelet-activating factor acetylhydrolase from human plasma. J Biol Chem. 1989 Apr 5;264(10):5331–5334. [PubMed]
  • Stremler KE, Stafforini DM, Prescott SM, McIntyre TM. Human plasma platelet-activating factor acetylhydrolase. Oxidatively fragmented phospholipids as substrates. J Biol Chem. 1991 Jun 15;266(17):11095–11103. [PubMed]
  • Stafforini DM, Prescott SM, McIntyre TM. Human plasma platelet-activating factor acetylhydrolase. Purification and properties. J Biol Chem. 1987 Mar 25;262(9):4223–4230. [PubMed]
  • Stafforini DM, McIntyre TM, Carter ME, Prescott SM. Human plasma platelet-activating factor acetylhydrolase. Association with lipoprotein particles and role in the degradation of platelet-activating factor. J Biol Chem. 1987 Mar 25;262(9):4215–4222. [PubMed]
  • Klimov AN, Nikiforova AA, Pleskov VM, Kuz'min AA, Kalashnikova NN. Zashchitnoe deistvie lipoproteidov vysokoi plotnosti, ikh podfraktsii i letsitin-kholesterin-atsiltransferazy v perekisnoi modifikatsii lipoproteidov nizkoi plotnosti. Biokhimiia. 1989 Jan;54(1):118–123. [PubMed]
  • Parthasarathy S, Barnett J, Fong LG. High-density lipoprotein inhibits the oxidative modification of low-density lipoprotein. Biochim Biophys Acta. 1990 May 22;1044(2):275–283. [PubMed]
  • Stafforini DM, Zimmerman GA, McIntyre TM, Prescott SM. The platelet-activating factor acetylhydrolase from human plasma prevents oxidative modification of low-density lipoprotein. Trans Assoc Am Physicians. 1992;105:44–63. [PubMed]
  • Watson AD, Navab M, Hama SY, Sevanian A, Prescott SM, Stafforini DM, McIntyre TM, Du BN, Fogelman AM, Berliner JA. Effect of platelet activating factor-acetylhydrolase on the formation and action of minimally oxidized low density lipoprotein. J Clin Invest. 1995 Feb;95(2):774–782. [PMC free article] [PubMed]
  • Chung BH, Wilkinson T, Geer JC, Segrest JP. Preparative and quantitative isolation of plasma lipoproteins: rapid, single discontinuous density gradient ultracentrifugation in a vertical rotor. J Lipid Res. 1980 Mar;21(3):284–291. [PubMed]
  • BLIGH EG, DYER WJ. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. [PubMed]
  • AMES BN, DUBIN DT. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed]
  • Kaluzny MA, Duncan LA, Merritt MV, Epps DE. Rapid separation of lipid classes in high yield and purity using bonded phase columns. J Lipid Res. 1985 Jan;26(1):135–140. [PubMed]
  • Zimmerman GA, McIntyre TM, Mehra M, Prescott SM. Endothelial cell-associated platelet-activating factor: a novel mechanism for signaling intercellular adhesion. J Cell Biol. 1990 Feb;110(2):529–540. [PMC free article] [PubMed]
  • Zimmerman GA, McIntyre TM, Prescott SM. Thrombin stimulates the adherence of neutrophils to human endothelial cells in vitro. J Clin Invest. 1985 Dec;76(6):2235–2246. [PMC free article] [PubMed]
  • Clay KL. Quantitation of platelet-activating factor by gas chromatography-mass spectrometry. Methods Enzymol. 1990;187:134–142. [PubMed]
  • Stremler KE, Stafforini DM, Prescott SM, McIntyre TM. Human plasma platelet-activating factor acetylhydrolase. Oxidatively fragmented phospholipids as substrates. J Biol Chem. 1991 Jun 15;266(17):11095–11103. [PubMed]
  • Tjoelker LW, Wilder C, Eberhardt C, Stafforini DM, Dietsch G, Schimpf B, Hooper S, Le Trong H, Cousens LS, Zimmerman GA, et al. Anti-inflammatory properties of a platelet-activating factor acetylhydrolase. Nature. 1995 Apr 6;374(6522):549–553. [PubMed]
  • Benveniste J, Nunez D, Duriez P, Korth R, Bidault J, Fruchart JC. Preformed PAF-acether and lyso PAF-acether are bound to blood lipoproteins. FEBS Lett. 1988 Jan 4;226(2):371–376. [PubMed]
  • Tanaka T, Minamino H, Unezaki S, Tsukatani H, Tokumura A. Formation of platelet-activating factor-like phospholipids by Fe2+/ascorbate/EDTA-induced lipid peroxidation. Biochim Biophys Acta. 1993 Feb 24;1166(2-3):264–274. [PubMed]
  • Dentan C, Lesnik P, Chapman MJ, Ninio E. PAF-acether-degrading acetylhydrolase in plasma LDL is inactivated by copper- and cell-mediated oxidation. Arterioscler Thromb. 1994 Mar;14(3):353–360. [PubMed]
  • Ambrosio G, Oriente A, Napoli C, Palumbo G, Chiariello P, Marone G, Condorelli M, Chiariello M, Triggiani M. Oxygen radicals inhibit human plasma acetylhydrolase, the enzyme that catabolizes platelet-activating factor. J Clin Invest. 1994 Jun;93(6):2408–2416. [PMC free article] [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem chemical compound records that cite the current articles. These references are taken from those provided on submitted PubChem chemical substance records. Multiple substance records may contribute to the PubChem compound record.
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem chemical substance records that cite the current articles. These references are taken from those provided on submitted PubChem chemical substance records.

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...