Research Article| Volume 240, ISSUE 1, P234-241, May 2015

Download started.


The nitroxide radical TEMPOL prevents obesity, hyperlipidaemia, elevation of inflammatory cytokines, and modulates atherosclerotic plaque composition in apoE−/− mice


      • Nitroxides scavenge radicals, are superoxide dismutase mimics and enzyme inhibitors.
      • Apo E−/− mice were fed a high fat diet without or with the nitroxide TEMPOL.
      • TEMPOL increased leptin and decreased obesity, adiponectin, plasma lipids and cytokines.
      • Plaque collagen and macrophage numbers were increased, and lipid levels decreased.
      • TEMPOL suppresses metabolic changes and increases atherosclerotic plaque stability.



      The nitroxide compound TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl radical) has been shown to prevent obesity-induced changes in adipokines in cell and animal systems. In this study we investigated whether supplementation with TEMPOL inhibits inflammation and atherosclerosis in apoE−/− mice fed a high fat diet (HFD).


      ApoE−/− mice were fed for 12 weeks on standard chow diet or a high-fat diet. Half the mice were supplemented with 10 mg/g TEMPOL in their food. Plasma samples were analysed for triglycerides, cholesterol, low- and high-density lipoprotein cholesterol, inflammatory cytokines and markers (interleukin-6, IL-6; monocyte-chemotactic protein, MCP-1; myeloperoxidase, MPO; serum amyloid A, SAA; adiponectin; leptin). Plaques in the aortic sinus were analysed for area, and content of collagen, lipid, macrophages and smooth muscle cells.


      High fat feeding resulted in marked increases in body mass and plasma lipid levels. Dietary TEMPOL decreased both parameters. In the high-fat-fed mice significant elevations in plasma lipid levels and the inflammatory markers IL-6, MCP-1, MPO, SAA were detected, along with an increase in leptin and a decrease in adiponectin. TEMPOL supplementation reversed these effects. When compared to HFD-fed mice, TEMPOL supplementation increased plaque collagen content, decreased lipid content and increased macrophage numbers.


      These data indicate that in a well-established model of obesity-associated hyperlipidaemia and atherosclerosis, TEMPOL had a significant impact on body mass, atherosclerosis, hyperlipidaemia and inflammation. TEMPOL may therefore be of value in suppressing obesity, metabolic disorders and increasing atherosclerotic plaque stability.

      Graphical abstract


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Graham I.
        • Cooney M.-T.
        • Bradley D.
        • et al.
        Dyslipidemias in the prevention of cardiovascular disease: risks and causality.
        Curr. Cardiol. Rep. 2012; 14: 709-720
        • Mitchell J.B.
        • Xavier S.
        • DeLuca A.M.
        • et al.
        A low molecular weight antioxidant decreases weight and lowers tumor incidence.
        Free Radic. Biol. Med. 2003; 34: 93-102
        • Soule B.P.
        • Hyodo F.
        • Matsumoto K.
        • et al.
        The chemistry and biology of nitroxide compounds.
        Free Radic. Biol. Med. 2007; 42: 1632-1650
        • Soule B.P.
        • Hyodo F.
        • Matsumoto K.
        • et al.
        Therapeutic and clinical applications of nitroxide compounds.
        Antioxid. Redox Signal. 2007; 9: 1731-1743
        • Samuni A.
        • Mitchell J.B.
        • DeGraff W.
        • et al.
        Nitroxide SOD-mimics: modes of action.
        Free Radic. Res. Commun. 1991; 12–13: 187-194
        • Krishna C.M.
        • Samuni A.
        Nitroxides as antioxidants.
        Meth. Enzymol. 1994; 234: 580-589
        • Lam M.A.
        • Pattison D.I.
        • Bottle S.E.
        • et al.
        Nitric oxide and nitroxides can act as efficient scavengers of protein-derived free radicals.
        Chem. Res. Toxicol. 2008; 21: 2111-2119
        • Pattison D.I.
        • Lam M.
        • Shinde S.S.
        • et al.
        The nitroxide TEMPO is an efficient scavenger of protein radicals: cellular and kinetic studies.
        Free Radic. Biol. Med. 2012; 53: 1664-1674
        • Krishna C.M.
        • Samuni A.
        • Taira J.
        • et al.
        Stimulation by nitroxides of catalase-like activity of heme proteins; kinetics and mechanism.
        J. Biol. Chem. 1996; 271: 26018-26025
        • Vaz S.M.
        • Augusto O.
        Inhibition of myeloperoxidase-mediated protein nitration by tempol: kinetics, mechanism, and implications.
        Proc. Natl. Acad. Sci. U. S. A. 2008; 105: 8191-8196
        • Rees M.D.
        • Bottle S.E.
        • Fairfull-Smith K.E.
        • et al.
        Inhibition of myeloperoxidase-mediated hypochlorous acid production by nitroxides.
        Biochem. J. 2009; 421: 79-86
        • Queiroz R.F.
        • Vaz S.M.
        • Augusto O.
        Inhibition of the chlorinating activity of myeloperoxidase by tempol: revisiting the kinetics and mechanisms.
        Biochem. J. 2011; 439: 423-431
        • Li F.
        • Jiang C.
        • Krausz K.W.
        • et al.
        Microbiome remodelling leads to inhibition of intestinal farnesoid X receptor signalling and decreased obesity.
        Nat. Commun. 2013; 4: 1-10
        • Frances D.E.
        • Ronco M.T.
        • Ingaramo P.I.
        • et al.
        Role of reactive oxygen species in the early stages of liver regeneration in streptozotocin-induced diabetic rats.
        Free Radic. Res. 2011; 45: 1143-1153
        • Wilcox C.S.
        • Pearlman A.
        Chemistry and antihypertensive effects of Tempol and other nitroxides.
        Pharmacol. Rev. 2008; 60: 418-469
        • Patel K.
        • Chen Y.F.
        • Dennehy K.
        • et al.
        Acute antihypertensive action of nitroxides in the spontaneously hypertensive rat.
        Am. J. Physiol. 2006; 290: R37-R43
        • Cannizzo B.
        • Quesada I.
        • Militello R.
        • et al.
        Tempol attenuates atherosclerosis associated with metabolic syndrome via decreased vascular inflammation and NADPH-2 oxidase expression.
        Free Radic. Res. 2014; 48: 684-693
        • Brown B.E.
        • Kim C.H.
        • Torpy F.R.
        • et al.
        Supplementation with carnosine decreases plasma triglycerides and modulates atherosclerotic plaque composition in diabetic apo E(-/-) mice.
        Atherosclerosis. 2014; 232: 403-409
        • Bursill C.A.
        • Choudhury R.P.
        • Ali Z.
        • et al.
        Broad-spectrum CC-chemokine blockade by gene transfer inhibits macrophage recruitment and atherosclerotic plaque formation in apolipoprotein E-knockout mice.
        Circulation. 2004; 110: 2460-2466
        • Allen J.K.
        • Hensley W.J.
        • Nicholls A.V.
        • et al.
        An enzymic and centrifugal method for estimating high-density lipoprotein cholesterol.
        Clin. Chem. 1979; 25: 325-327
        • Di Bartolo B.A.
        • Vanags L.Z.
        • Tan J.T.
        • et al.
        The apolipoprotein A-I mimetic peptide, ETC-642, reduces chronic vascular inflammation in the rabbit.
        Lipids Health Dis. 2011; 10: 224
        • Ankel E.G.
        • Lai C.S.
        • Hopwood L.E.
        • et al.
        Cytotoxicity of commonly used nitroxide radical spin probes.
        Life Sci. 1987; 40: 495-498
        • Mitchell J.B.
        • Anver M.R.
        • Sowers A.L.
        • et al.
        The antioxidant Tempol reduces carcinogenesis and enhances survival in mice when administered after nonlethal total body radiation.
        Cancer Res. 2012; 72: 4846-4855
        • Finn A.V.
        • Nakano M.
        • Narula J.
        • et al.
        Concept of vulnerable/unstable plaque.
        Arterioscler. Thromb. Vasc. Biol. 2010; 30: 1282-1292
        • Dandona P.
        • Aljada A.
        • Chaudhuri A.
        • et al.
        Metabolic syndrome - a comprehensive perspective based on interactions between obesity, diabetes, and inflammation.
        Circulation. 2005; 111: 1448-1454
        • Hotamisligil G.S.
        • Shargill N.S.
        • Spiegelman B.M.
        Adipose expression of tumor-necrosis-factor-alpha - direct role in obesity-linked insulin resistance.
        Science. 1993; 259: 87-91
        • Knight S.F.
        • Yuan J.H.
        • Roy S.
        • et al.
        Simvastatin and tempol protect against endothelial dysfunction and renal injury in a model of obesity and hypertension.
        Am. J. Physiol. 2010; 298: F86-F94
        • Virdis A.
        • Santini F.
        • Colucci R.
        • et al.
        Vascular generation of tumor necrosis factor-alpha reduces nitric oxide availability in small arteries from visceral fat of obese patients.
        J. Am. Coll. Cardiol. 2011; 58: 238-247
        • Queiroz R.F.
        • Jordao A.K.
        • Cunha A.C.
        • et al.
        Nitroxides attenuate carrageenan-induced inflammation in rat paws by reducing neutrophil infiltration and the resulting myeloperoxidase-mediated damage.
        Free Radic. Biol. Med. 2012; 53: 1942-1953
        • Baetta R.
        • Corsini A.
        Role of polymorphonuclear neutrophils in atherosclerosis: current state and future perspectives.
        Atherosclerosis. 2010; 210: 1-13
        • van Leeuwen M.
        • Gijbels M.J.
        • Duijvestijn A.
        • et al.
        Accumulation of myeloperoxidase-positive neutrophils in atherosclerotic lesions in LDLR-/- mice.
        Arterioscler. Thromb. Vasc. Biol. 2008; 28: 84-89
        • Daugherty A.
        • Dunn J.L.
        • Rateri D.L.
        • et al.
        Myeloperoxidase, a catalyst for lipoprotein oxidation, is expressed in human atherosclerotic lesions.
        J. Clin. Invest. 1994; 94: 437-444
        • Sugiyama S.
        • Okada Y.
        • Sukhova G.K.
        • et al.
        Macrophage myeloperoxidase regulation by granulocyte macrophage colony- stimulating factor in human atherosclerosis and implications in acute coronary syndromes.
        Am. J. Pathol. 2001; 158: 879-891
        • Tang W.H.
        • Wu Y.
        • Nicholls S.J.
        • et al.
        Plasma myeloperoxidase predicts incident cardiovascular risks in stable patients undergoing medical management for coronary artery disease.
        Clin. Chem. 2011; 57: 33-39
        • Nicholls S.J.
        • Hazen S.L.
        Myeloperoxidase and cardiovascular disease.
        Arterioscler. Thromb. Vasc. Biol. 2005; 25: 1102-1111
        • Kajer T.B.
        • Fairfull-Smith K.E.
        • Y T.
        • et al.
        Inhibition of myeloperoxidase- and neutrophil-mediated oxidant production by tetraethyl- and tetramethyl-nitroxides.
        Free Radic. Biol. Med. 2014; 70: 96-105
        • Small D.M.
        • Shipley G.G.
        Physical-chemical basis of lipid deposition in atherosclerosis.
        Science. 1974; 185: 222-229
        • Libby P.
        Inflammation in atherosclerosis.
        Nature. 2002; 420: 868-874
        • Menini S.
        • Iacobini C.
        • Ricci C.
        • et al.
        D-carnosine octylester attenuates atherosclerosis and renal disease in ApoE null mice fed a western diet through reduction of carbonyl stress and inflammation.
        Br. J. Pharmacol. 2012; 166: 1344-1356
        • Ebenezer P.J.
        • Mariappan N.
        • Elks C.M.
        • et al.
        Diet-induced renal changes in Zucker rats are ameliorated by the superoxide dismutase mimetic TEMPOL.
        Obesity. 2009; 17: 1994-2002
        • Asghar M.
        • Lokhandwala M.F.
        Antioxidant Tempol lowers age-related increases in insulin resistance in Fischer 344 rats.
        Clin. Exp. Hypertens. 2006; 28: 533-541
        • Jernas M.
        • Palming J.
        • Sjoholm K.
        • et al.
        Separation of human adipocytes by size: hypertrophic fat cells display distinct gene expression.
        Faseb J. 2006; 20: 1540-1542
        • Kumon Y.
        • Suehiro T.
        • Hashimoto K.
        • et al.
        Local expression of acute phase serum amyloid A mRNA in rheumatoid arthritis synovial tissue and cells.
        J. Rheumatol. 1999; 26: 785-790
        • Poitou C.
        • Viguerie N.
        • Cancello R.
        • et al.
        Serum amyloid A: production by human white adipocyte and regulation by obesity and nutrition.
        Diabetologia. 2005; 48: 519-528
        • Meek R.L.
        • Urielishoval S.
        • Benditt E.P.
        Expression of apolipoprotein serum amyloid-a messenger-rna in human atherosclerotic lesions and cultured vascular cells – implications for serum amyloid-A function.
        Proc. Natl. Acad. Sci. U. S. A. 1994; 91: 3186-3190
        • Hattori Y.
        • Akimoto K.
        • Gross S.S.
        • et al.
        Angiotensin-II-induced oxidative stress elicits hypoadiponectinaemia rats.
        Diabetologia. 2005; 48: 1066-1074
        • Villarreal-Molina M.T.
        • Antuna-Puente B.
        Adiponectin: anti-inflammatory and cardioprotective effects.
        Biochimie. 2012; 94: 2143-2149
        • Al-Daghri N.M.
        • Al-Attas O.S.
        • Alokail M.S.
        • et al.
        Visceral adiposity index is highly associated with adiponectin values and glycaemic disturbances.
        Eur. J. Clin. Invest. 2013; 43: 183-189
        • Enos R.T.
        • Davis J.M.
        • Velazquez K.T.
        • et al.
        Influence of dietary saturated fat content on adiposity, macrophage behavior, inflammation, and metabolism: composition matters.
        J. Lipid Res. 2013; 54: 152-163
        • Samuni Y.
        • Cook J.A.
        • Choudhuri R.
        • et al.
        Inhibition of adipogenesis by Tempol in 3T3-L1 cells.
        Free Radic. Biol. Med. 2010; 49: 667-673