Macrophage NADPH oxidase activation, impaired cholesterol fluxes, and increased cholesterol biosynthesis in diabetic mice: A stimulatory role for d-glucose

  • Author Footnotes
    1 These authors contributed equally to this work.
    Tony Hayek
    Corresponding author at: Internal Medicine Department E, Rambam Medical Center, 31096 Haifa, Israel. Tel.: +972 4 854 2059; fax: +972 4 854 2359.
    1 These authors contributed equally to this work.
    Lipid research Laboratory, Faculty of Medicine, Technion, Rambam Medical Center, Haifa, Israel
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  • Author Footnotes
    1 These authors contributed equally to this work.
    Marielle Kaplan
    1 These authors contributed equally to this work.
    Clinical Biochemistry Department, Rambam Medical Center, Haifa, Israel
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  • Rachel Kerry
    Lipid research Laboratory, Faculty of Medicine, Technion, Rambam Medical Center, Haifa, Israel
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  • Michael Aviram
    Lipid research Laboratory, Faculty of Medicine, Technion, Rambam Medical Center, Haifa, Israel
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  • Author Footnotes
    1 These authors contributed equally to this work.


      Diabetes is clearly associated with accelerated atherosclerosis development, but molecular mechanisms involved in diabetes-induced atherosclerosis remain to be clarified. The aim of this study was to identify cellular mechanisms involved in diabetes-induced macrophage foam cell formation, the hallmark of early atherogenesis. Mouse peritoneal macrophages (MPMs) isolated from Balb-C streptozotocin-induced diabetic mice, exhibited significantly higher total peroxides, lipid peroxides and paraoxonase 2 (PON2) activity by 290%, 61% and 55%, respectively, compared to non-diabetic mice. In vitro studies revealed that glucose-induced oxidative stress was obtained by d-glucose, but not by l-glucose and it involved activation of the NADPH oxidase complex, and up-regulation of the macrophage PON2.
      Next, MPMs isolated from Balb-C diabetic mice, compared to control Balb-C mice, demonstrated increased cholesterol content by 4.2-fold associated with increased cholesterol biosynthesis and increased uptake of oxidized LDL (Ox-LDL) by 5.9-fold and 31%, respectively. These effects on cellular cholesterol metabolism were associated with up-regulation of the scavenger receptors for Ox-LDL (CD-36 and SR-A), and of HMG-CoA reductase (cholesterol biosynthesis rate limiting enzyme). Finally, using pravastatin (inhibitor of HMG-CoA reductase) and the antioxidant Vitamin E, we have shown that d-glucose-induced macrophage oxidative stress is secondary to its stimulatory effect on macrophage cholesterol biosynthesis.
      In conclusion, macrophages from diabetic mice demonstrate increased oxidative stress associated with activation of NADPH oxidase and up-regulation of cellular PON2, as well as increased macrophages cholesterol uptake and biosynthesis (increased expression of CD-36 and HMG-CoA reductase). The above mechanisms in diabetic mice could be the result of the effect of high d-glucose on macrophages.


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        • Garcia M.J.
        • McNamara P.M.
        • Gordon T.
        • Kannel W.B.
        Morbidity and mortality in diabetics in the Framingham population: sixteen year follow-up study.
        Diabetes. 1974; 23: 105-111
        • Uusitopa M.I.
        • Niskanen L.K.
        • Siitonen O.
        • Voutilainen E.
        • Pyorala K.
        Five-year incidence of atherosclerotic vascular disease in relation to general risk factors, insulin level, and abnormalities in lipoprotein composition in non-insulin-dependent diabetic and nondiabetic subjects.
        Circulation. 1990; 82: 27-36
        • Lusis A.J.
        Nature. 2000; 407: 233-241
        • Aviram M.
        Review of human studies on oxidative damage and antioxidant protection related to cardiovascular diseases.
        Free Radic Res. 2000; 33: S85-S97
        • Steinberg D.
        • Lewis A.
        Oxidative modification of LDL and atherogenesis.
        Circulation. 1997; 95: 1062-1071
        • Berliner J.A.
        • Navab M.
        • Fogelman A.M.
        • et al.
        Atherosclerosis: Basic mechanisms: oxidation inflammation and genetics.
        Circulation. 1995; 91: 2488-2496
        • Aviram M.
        • Hardak E.
        • Vaya J.
        • et al.
        Human serum paraoxonases Q and R selectively decrease lipid peroxides in human coronary and carotid atherosclerotic lesions: PON1 esterase and peroxidase-like activities.
        Circulation. 2000; 30: 2510-2517
        • Rosenblat M.
        • Draganov D.
        • Watson C.E.
        • et al.
        Mouse macrophage paraoxonase 2 activity is increased whereas cellular paraoxonase 3 activity is decreased under oxidative stress.
        Arterioscler Thromb Vasc Biol. 2003; 23: 468-474
        • Rosenblat M.
        • Aviram M.
        Nutritional pharmacological and metabolic influences on paraoxonase.
        Curr Opin Lipidol. 2005; 16: 393-399
        • Aviram M.
        • Rosenblat M.
        Paraoxonases and cardiovascular diseases: pharmacological and nutritional influences.
        Curr Opin Lipidol. 2005; 16: 393-399
        • Maziere C.
        • Auclair M.
        • Rose-Robert F.
        • Leflon P.
        • Maziere J.C.
        Glucose-enriched medium enhances cell-mediated low density lipoprotein peroxidation.
        FEBS Lett. 1995; 363: 277-279
        • Griffin E.
        • Re A.
        • Hamel N.
        • et al.
        A link between diabetes and atherosclerosis: glucose regulates expression of CD36 at the level of translation.
        Nat Med. 2001; 7: 840-846
        • Hayek T.
        • Hussein K.
        • Aviram M.
        • et al.
        Macrophage-foam cell formation in streptozotocin-induced diabetic mice: stimulatory effect of glucose.
        Atherosclerosis. 2005; 183: 25-33
        • Aviram M.
        • Rosenblat M.
        • Bisgaier C.L.
        • et al.
        Paraoxonase inhibits high-density lipoprotein oxidation and preserves its functions. A possible peroxidative role for paraoxonase.
        J Clin Invest. 1998; 101: 1581-1590
        • Royall J.A.
        • Ischiropoulos H.
        Evaluation of 2′,7′-dichlorofluorescin and dihydrorhodamine 123 as fluorescent probes for intracellular H2O2 in cultured endothelial cells.
        Arch Biochem Biophys. 1993; 302: 348-355
        • Maor I.
        • Kaplan M.
        • Hayek T.
        • et al.
        Oxidized monocyte-derived macrophages in aortic atherosclerotic lesion from E0 mice and from human carotid artery contain lipid peroxides and oxysterols.
        Biochem Biophys Res Commun. 2000; 269: 775-780
        • Aviram M.
        • Vaya J.
        Markers for LDL oxidation.
        Methods Enzymol. 2001; 335: 244-256
        • Shamir R.
        • Hartman C.
        • Karry R.
        • et al.
        Paraoxonases (PONs) 1, 2, 3 are expressed in human and in mice gastrointestinal tract and in human intestinal Caco2cell line: selective secretion of PON1 and PON2.
        Free Radic Biol Res. 2005; 39: 336-344
        • Johnston R.B.
        • Keele Jr., B.B.
        • Mirsa H.P.
        • et al.
        The role of superoxide anion generation in phagocytic bactericidal activity.
        J Clin Invest. 1975; 55: 1357-1372
        • Kaplan M.
        • Aviram M.
        • Knopf C.
        • Keidar S.
        Angiotensin II reduces macrophage cholesterol efflux: a role for the AT-1 receptor but not for the ABC1 transporter.
        Biochem Biophys Res Commun. 2002; 290: 1529-1534
        • Fuhrman B.
        • Volkova N.
        • Aviram M.
        Pomegranate juice inhibits oxidized LDL uptake and cholesterol biosynthesis in macrophages.
        J Nutr Biochem. 2005; 16: 570-576
        • Rozenberg O.
        • Shih D.M.
        • Aviram M.
        Human serum paraoxonase decreases macrophage cholesterol biosynthesis: a possible role for its phospholipase A2-like activity and lysophosphatidylcholine formation.
        Arterioscler Thromb Vasc Biol. 2003; 23: 461-467
        • Han J.
        • Hajjar D.P.
        • Febbraio M.
        • Nicholson A.C.
        Native and modified low density lipoproteins increase the functional expression of the macrophage class B scavenger receptor, CD36.
        J Biol Chem. 1997; 272: 21654-21659
        • Mietus-Snyder M.
        • Gowri M.S.
        • Pitas R.E.
        Class A scavenger receptor up-regulation in smooth muscle cells by oxidized low density lipoprotein. Enhancement by calcium flux and concurrent cyclooxygenase-2 up-regulation.
        J Biol Chem. 2000; 275: 17661-17670
        • Iida K.T.
        • Kawakami Y.
        • Suzuki H.
        • et al.
        PPAR gamma ligands, troglitazone and pioglitazone, up-regulate expression of HMG-CoA synthase and HMG-CoA reductase gene in THP-1 macrophages.
        FEBS Lett. 2002; 520: 177-181
        • Rosenblat M.
        • Aviram M.
        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. 2002; 160: 69-80
        • Cathcart M.K.
        Regulation of superoxide anion production by NADPH oxidase in monocytes/macrophages: contributions to atherosclerosis.
        Arterioscler Thromb Vasc Biol. 2004; 24 (2003): 23-28
        • Mackness B.
        • McElduff P.
        • Mackness M.I.
        J Intern Med. 2005; 258: 363-368
        • Rosenblat M.
        • Karry R.
        • Aviram M.
        Paraoxonase 1 (PON1) is a more potent antioxidant and stimulant of macrophage cholesterol efflux, when present in HDL than in lipoprotein-deficient serum: relevance to diabetes.
        Atherosclerosis. 2006; 187: 74-81
        • Lagor William R.
        • de Groh Eric D.
        • Nes Gene C.
        Diabetes alters the occupancy of the hepatic 3-hydroxy-3-methylglutaryl-CoA reductase promoter.
        J Biol Chem. 2005; 280: 36601-36608
        • Rosenson R.S.
        Pluripotential mechanisms of cardioprotection with HMG-CoA reductase inhibitor therapy.
        Am J Cardiovasc Drugs. 2001; 1: 411-420
        • Fukuhara-Takaki K.
        • Sakai M.
        • Sakamoto Y.
        • Takeya M.
        • Horiuchi S.
        Expression of class A scavenger receptor is enhanced by high glucose in vitro and under diabetic conditions in vivo: one mechanism for an increased rate of atherosclerosis in diabetes.
        J Biol Chem. 2005; 280: 3355-3364