Skip to main content
    • Aa
    • Aa

Differential effects of apolipoprotein E3 and E4 on markers of oxidative status in macrophages

  • Laia Jofre-Monseny (a1), Sonia de Pascual-Teresa (a2), Eva Plonka (a1), Patricia Huebbe (a1), Christine Boesch-Saadatmandi (a1), Anne-Marie Minihane (a3) and Gerald Rimbach (a1)...

ApoE is secreted by macrophages at the lesion site of the atherosclerotic plaque, where it is thought to play a protective role against atherosclerosis independently of its effects on lipid metabolism. Of the three common isoforms for apoE, apoE4 is associated with higher risk of cardiovascular disease (CVD). In vitro studies have shown that recombinant apoE may act as an antioxidant in an isoform-dependent manner (E2>E3>E4). The oxidative status of the macrophages plays a key role in the process of atherosclerosis. In the present study the possible differential actions of apoE3 and apoE4 on several parameters of oxidative status were determined in stably transfected murine macrophages (RAW 264·7-apoE3 and -apoE4). No differences between genotypes were observed after peroxide challenge in either protection against cytotoxicity or in cell membrane oxidation, and modest differences were observed in the non-enzymatic antioxidants (glutathione and α-tocopherol) in apoE3 v. apoE4 macrophages. Importantly, cells secreting apoE4 showed increased membrane oxidation under basal conditions, and produced more NO and superoxide anion radicals than the apoE3 macrophages after stimulation. The present data suggest that apoE genotype influences the oxidative status of macrophages, and this could partly contribute to the higher CVD risk observed in apoE4 carriers.

  • View HTML
    • Send article to Kindle

      To send this article to your Kindle, first ensure is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about sending to your Kindle.

      Note you can select to send to either the or variations. ‘’ emails are free but can only be sent to your device when it is connected to wi-fi. ‘’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

      Find out more about the Kindle Personal Document Service.

      Differential effects of apolipoprotein E3 and E4 on markers of oxidative status in macrophages
      Available formats
      Send article to Dropbox

      To send this article to your Dropbox account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your Dropbox account. Find out more about sending content to Dropbox.

      Differential effects of apolipoprotein E3 and E4 on markers of oxidative status in macrophages
      Available formats
      Send article to Google Drive

      To send this article to your Google Drive account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your Google Drive account. Find out more about sending content to Google Drive.

      Differential effects of apolipoprotein E3 and E4 on markers of oxidative status in macrophages
      Available formats
Corresponding author
*Corresponding author: Prof. G. Rimbach, fax +49 0431 88802628, email
Hide All
Ali K, Middleton M, Pure E & Rader DJ (2005) Apolipoprotein E suppresses the type I inflammatory response in vivo. Circ Res 97, 922927.
Asehnoune K, Strassheim D, Mitra S, Kim JY & Abraham E (2004) Involvement of reactive oxygen species in Toll-like receptor 4-dependent activation of NF-kappa B. J Immunol 172, 25222529.
Aviram M & Fuhrman B (1998) LDL oxidation by arterial wall macrophages depends on the oxidative status in the lipoprotein and in the cells: role of prooxidants vs. antioxidants. Mol Cell Biochem 188, 149159.
Beckman JS & Koppenol WH (1996) Nitric oxide, superoxide, and peroxynitrite: the good, the bad, and ugly. Am J Physiol 271, C1424C1437.
Bellosta S, Mahley RW, Sanan DA, Murata J, Newland DL, Taylor JM & Pitas RE (1995) Macrophage-specific expression of human apolipoprotein E reduces atherosclerosis in hypercholesterolemic apolipoprotein E-null mice. J Clin Invest 96, 21702179.
Bokoch GM & Knaus UG (2003) NADPH oxidases: not just for leukocytes anymore! Trends Biochem Sci 28, 502508.
Borenfreund E & Puerner JA (1985) Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicol Lett 24, 119124.
Choi HS, Kim JW, Cha YN & Kim C (2006) A quantitative nitroblue tetrazodium assay for determining intracellular superoxide anion production in phagocytic cells. J Immunoassay Immunochem 27, 3144.
Colton CA, Brown CM, Cook D, Needham LK, Xu Q, Czapiga M, Saunders AM, Schmechel DE, Rasheed K & Vitek MP (2002) APOE and the regulation of microglial nitric oxide production: a link between genetic risk and oxidative stress. Neurobiol Aging 23, 777785.
Colton CA, Czapiga M, Snell-Callanan J, Chernyshev ON & Vitek MP (2001) Apolipoprotein E acts to increase nitric oxide production in macrophages by stimulating arginine transport. Biochim Biophys Acta 1535, 134144.
Colton CA, Needham LK, Brown C, Cook D, Rasheed K, Burke JR, Strittmatter WJ, Schmechel DE & Vitek MP (2004) APOE genotype-specific differences in human and mouse macrophage nitric oxide production. J Neuroimmunol 147, 6267.
Dietrich M, Hua Y, Block G, Olano E, Packer L, Morrow JD, Hudes M, Abdukeyum G, Rimbach G & Minihane AM (2005) Associations between apolipoprotein E genotype and circulating F2-isoprostane levels in humans. Lipids 40, 329334.
Drummen GP, Makkinje M, Verkleij AJ, Op den Kamp JA & Post JA (2004) Attenuation of lipid peroxidation by antioxidants in rat-1 fibroblasts: comparison of the lipid peroxidation reporter molecules cis-parinaric acid and C11-BODIPY(581/591) in a biological setting. Biochim Biophys Acta 1636, 136150.
Duan H, Lin CY & Mazzone T (1997) Degradation of macrophage ApoE in a nonlysosomal compartment. Regulation by sterols. J Biol Chem 272, 3115631162.
Fazio S, Babaev VR, Murray AB, Hasty AH, Carter KJ, Gleaves LA, Atkinson JB & Linton MF (1997) Increased atherosclerosis in mice reconstituted with apolipoprotein E null macrophages. Proc Natl Acad Sci U S A 94, 46474652.
Fernandes MA, Proenca MT, Nogueira AJ, Grazina MM, Oliveira LM, Fernandes AI, Santiago B, Santana I & Oliveira CR (1999) Influence of apolipoprotein E genotype on blood redox status of Alzheimer's disease patients. Int J Mol Med 4, 179186.
Gao R, Stone WL, Huang T, Papas AM & Qui M (2002) The uptake of tocopherols by RAW 264·7 macrophages. Nutr J 1, 2.
Gotoh N, Graham A, Nikl E & Darley-Usmar VM (1993) Inhibition of glutathione synthesis increases the toxicity of oxidized low-density lipoprotein to human monocytes and macrophages. Biochem J 296, Part 1, 151154.
Hara M, Matsushima T, Satoh H, Iso-o N, Noto H, Togo M, Kimura S, Hashimoto Y & Tsukamoto K (2003) Isoform-dependent cholesterol efflux from macrophages by apolipoprotein E is modulated by cell surface proteoglycans. Arterioscler Thromb Vasc Biol 23, 269274.
Hasty AH, Linton MF, Brandt SJ, Babaev VR, Gleaves LA & Fazio S (1999) Retroviral gene therapy in ApoE-deficient mice: ApoE expression in the artery wall reduces early foam cell lesion formation. Circulation 99, 25712576.
Hatters DM, Peters-Libeu CA & Weisgraber KH (2006) Apolipoprotein E structure: insights into function. Trends Biochem Sci 31, 445–454.
Hayek T, Oiknine J, Brook JG & Aviram M (1994) Increased plasma and lipoprotein lipid peroxidation in apo E-deficient mice. Biochem Biophys Res Commun 201, 15671574.
Hernandez-Montes E, Pollard SE, Vauzour D, Jofre-Montseny L, Rota C, Rimbach G, Weinberg PD & Spencer JP (2006) Activation of glutathione peroxidase via Nrf1 mediates genistein's protection against oxidative endothelial cell injury. Biochem Biophys Res Commun 346, 851–859.
Hixson JE & Vernier DT (1990) Restriction isotyping of human apolipoprotein E by gene amplification and cleavage with HhaI. J Lipid Res 31, 545–548.
Humphries SE, Talmud PJ, Hawe E, Bolla M, Day IN & Miller GJ (2001) Apolipoprotein E4 and coronary heart disease in middle-aged men who smoke: a prospective study. Lancet 358, 115–119.
Jialal I, Fuller CJ & Huet BA (1995) The effect of alpha-tocopherol supplementation on LDL oxidation. A dose-response study. Arterioscler Thromb Vasc Biol 15, 190198.
Jolivalt C, Leininger-Muller B, Bertrand P, Herber R, Christen Y & Siest G (2000) Differential oxidation of apolipoprotein E isoforms and interaction with phospholipids. Free Radic Biol Med 28, 129–140.
Kelly ME, Clay MA, Mistry MJ, Hsieh-Li HM & Harmony JA (1994) Apolipoprotein E inhibition of proliferation of mitogen-activated T lymphocytes: production of interleukin 2 with reduced biological activity. Cell Immunol 159, 124139.
Kitagawa K, Matsumoto M, Kuwabara K, Takasawa K, Tanaka S, Sasaki T, Matsushita K, Ohtsuki T, Yanagihara T & Hori M (2002) Protective effect of apolipoprotein E against ischemic neuronal injury is mediated through antioxidant action. J Neurosci Res 68, 226232.
Lodge JK, Hall WL, Jeanes YM & Proteggente AR (2004) Physiological factors influencing vitamin E biokinetics. Ann N Y Acad Sci 1031, 60–73.
Mardones P, Strobel P, Miranda S, Leighton F, Quinones V, Amigo L, Rozowski J, Krieger M & Rigotti A (2002) Alpha-tocopherol metabolism is abnormal in scavenger receptor class B type I (SR-BI)-deficient mice. J Nutr 132, 443449.
McCormick CC & Parker RS (2004) The cytotoxicity of vitamin E is both vitamer- and cell-specific and involves a selectable trait. J Nutr 134, 33353342.
Minihane AM, Khan S, Leigh-Firbank EC, Talmud P, Wright JW, Murphy MC, Griffin BA & Williams CM (2000) ApoE polymorphism and fish oil supplementation in subjects with an atherogenic lipoprotein phenotype. Arterioscler Thromb Vasc Biol 20, 1990–1997.
Mistry MJ, Clay MA, Kelly ME, Steiner MA & Harmony JA (1995) Apolipoprotein E restricts interleukin-dependent T lymphocyte proliferation at the G1A/G1B boundary. Cell Immunol 160, 14–23.
Miyata M & Smith JD (1996) Apolipoprotein E allele-specific antioxidant activity and effects on cytotoxicity by oxidative insults and beta-amyloid peptides. Nat Genet 14, 5561.
Naguib YM (1998) A fluorometric method for measurement of peroxyl radical scavenging activities of lipophilic antioxidants. Anal Biochem 265, 290298.
Pap EH, Drummen GP, Winter VJ, Kooij TW, Rijken P, Wirtz KW, Op den Kamp JA, Hage WJ & Post JA (1999) Ratio-fluorescence microscopy of lipid oxidation in living cells using C11-BODIPY(581/591). FEBS Lett 453, 278–282.
Pepe MG & Curtiss LK (1986) Apolipoprotein E is a biologically active constituent of the normal immunoregulatory lipoprotein, LDL-In. J Immunol 136, 37163723.
Peroutka SJ & Dreon DM (2000) The value of genotyping for apolipoprotein E alleles in relation to vitamin E supplementation. Eur J Pharmacol 410, 161–163.
Pham T, Kodvawala A & Hui DY (2005) The receptor binding domain of apolipoprotein E is responsible for its antioxidant activity. Biochemistry 44, 75777582.
Pratico D, Lee VM-Y, Trojanowski JQ, Rokach J & Fitzgerald GA (1998) Increased F2-isoprostanes in Alzheimer's disease: evidence for enhanced lipid peroxidation in vivo. FASEB J 12, 1777–1783.
Ramassamy C, Averill D, Beffert U, Theroux L, Lussier-Cacan S, Cohn JS, Christen Y, Schoofs A, Davignon J & Poirier J (2000) Oxidative insults are associated with apolipoprotein E genotype in Alzheimer's disease brain. Neurobiol Dis 7, 23–37.
Riddell DR, Graham A & Owen JS (1997) Apolipoprotein E inhibits platelet aggregation through the l-arginine:nitric oxide pathway. Implications for vascular disease. J Biol Chem 272, 8995.
Rimbach G, De Pascual-Teresa S, Ewins BA, Matsugo S, Uchida Y, Minihane AM, Turner R, VafeiAdou K & Weinberg PD (2003) Antioxidant and free radical scavenging activity of isoflavone metabolites. Xenobiotica 33, 913925.
Rosenblat M & Aviram M (2002) Oxysterol-induced activation of macrophage NADPH-oxidase enhances cell-mediated oxidation of LDL in the atherosclerotic apolipoprotein E deficient mouse: inhibitory role for vitamin E. Atherosclerosis 160, 69–80.
Rosenblat M, Coleman R & Aviram M (2002) Increased macrophage glutathione content reduces cell-mediated oxidation of LDL and atherosclerosis in apolipoprotein E-deficient mice. Atherosclerosis 163, 17–28.
Shimano H, Ohsuga J, Shimada M, Namba Y, Gotoda T, Harada K, Katsuki M, Yazaki Y & Yamada N (1995) Inhibition of diet-induced atheroma formation in transgenic mice expressing apolipoprotein E in the arterial wall. J Clin Invest 95, 469476.
Smith JD, Miyata M, Ginsberg M, Grigaux C, Shmookler E & Plump AS (1996) Cyclic AMP induces apolipoprotein E binding activity and promotes cholesterol efflux from a macrophage cell line to apolipoprotein acceptors. J Biol Chem 271, 3064730655.
Song Y, Stampfer MJ & Liu S (2004) Meta-analysis: apolipoprotein E genotypes and risk for coronary heart disease. Ann Intern Med 141, 137–147.
Stannard AK, Riddell DR, Sacre SM, Tagalakis AD, Langer C, von Eckardstein A, Cullen P, Athanasopoulos T, Dickson G & Owen JS (2001) Cell-derived apolipoprotein E (ApoE) particles inhibit vascular cell adhesion molecule-1 (VCAM-1) expression in human endothelial cells. J Biol Chem 276, 4601146016.
Talmud PJ, Stephens JW, Hawe E, Demissie S, Cupples LA, Hurel SJ, Humphries SE & Ordovas JM (2005) The significant increase in cardiovascular disease risk in APOEɛ4 carriers is evident only in men who smoke: potential relationship between reduced antioxidant status and ApoE4. Ann Hum Genet 69, 613–622.
Tamaoka A, Miyatake F, Matsuno S, et al. (2000) Apolipoprotein E allele-dependent antioxidant activity in brains with Alzheimer's disease. Neurology 54, 23192321.
Wang-Iverson P, Gibson JC & Brown WV (1985) Plasma apolipoprotein secretion by human monocyte-derived macrophages. Biochim Biophys Acta 834, 256–262.
Yamada N, Inoue I, Kawamura M, et al. (1992) Apolipoprotein E prevents the progression of atherosclerosis in Watanabe heritable hyperlipidemic rabbits. J Clin Invest 89, 706711.
Yoshida H, Hasty AH, Major AS, Ishiguro H, Su YR, Gleaves LA, Babaev VR, Linton MF & Fazio S (2001) Isoform-specific effects of apolipoprotein E on atherogenesis: gene transduction studies in mice. Circulation 104, 28202825.
Yoshida T (1996) Determination of reduced and oxidized glutathione in erythrocytes by high-performance liquid chromatography with ultraviolet absorbance detection. J Chromatogr B Biomed Appl 678, 157–164.
Zeleny M, Swertfeger DK, Weisgraber KH & Hui DY (2002) Distinct apolipoprotein E isoform preference for inhibition of smooth muscle cell migration and proliferation. Biochemistry 41, 1182011823.
Zhou J, Struthers AD & Lyles GA (1999) Differential effects of some cell signalling inhibitors upon nitric oxide synthase expression and nuclear factor-kappaB activation induced by lipopolysaccharide in rat aortic smooth muscle cells. Pharmacol Res 39, 363–373.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

British Journal of Nutrition
  • ISSN: 0007-1145
  • EISSN: 1475-2662
  • URL: /core/journals/british-journal-of-nutrition
Please enter your name
Please enter a valid email address
Who would you like to send this to? *



Full text views

Total number of HTML views: 2
Total number of PDF views: 58 *
Loading metrics...

Abstract views

Total abstract views: 134 *
Loading metrics...

* Views captured on Cambridge Core between September 2016 - 21st October 2017. This data will be updated every 24 hours.