Anti-oxidative effects of pomegranate juice (PJ) consumption by diabetic patients on serum and on macrophages

  • Mira Rosenblat
    The Lipid Research Laboratory, Technion Faculty of Medicine, The Rappaport Family Institute for Research in the Medical Sciences, Rambam Medical Center, 31096 Haifa, Israel
    Search for articles by this author
  • Tony Hayek
    The Lipid Research Laboratory, Technion Faculty of Medicine, The Rappaport Family Institute for Research in the Medical Sciences, Rambam Medical Center, 31096 Haifa, Israel

    Internal Medicine Department E, Rambam Medical Center, Haifa, Israel
    Search for articles by this author
  • Michael Aviram
    Corresponding author. Tel.: +972 4 8542970; fax: +972 4 8542130.
    The Lipid Research Laboratory, Technion Faculty of Medicine, The Rappaport Family Institute for Research in the Medical Sciences, Rambam Medical Center, 31096 Haifa, Israel
    Search for articles by this author


      Diabetes is associated with increased oxidative stress and atherosclerosis development. In the present study, we investigated the effects of pomegranate juice (PJ; which contains sugars and potent anti-oxidants) consumption by diabetic patients on blood diabetic parameters, and on oxidative stress in their serum and macrophages. Ten healthy subjects (controls) and 10 non-insulin dependent diabetes mellitus (NIDDM) patients who consumed PJ (50 ml per day for 3 months) participated in the study. In the patients versus controls serum levels of lipid peroxides and thiobarbituric acid reactive substances (TBARS) were both increased, by 350% and 51%, respectively, whereas serum SH groups content and paraoxonase 1 (PON1) activity, were both decreased (by 23%). PJ consumption did not affect serum glucose, cholesterol and triglyceride levels, but it resulted in a significant reduction in serum lipid peroxides and TBARS levels by 56% and 28%, whereas serum SH groups and PON1 activity significantly increased by 12% and 24%, respectively. In the patients versus controls monocytes-derived macrophages (HMDM), we observed increased level of cellular peroxides (by 36%), and decreased glutathione content (by 64%). PJ consumption significantly reduced cellular peroxides (by 71%), and increased glutathione levels (by 141%) in the patients’ HMDM. The patients’ versus control HMDM took up oxidized LDL (Ox-LDL) at enhanced rate (by 37%) and PJ consumption significantly decreased the extent of Ox-LDL cellular uptake (by 39%). We thus conclude that PJ consumption by diabetic patients did not worsen the diabetic parameters, but rather resulted in anti-oxidative effects on serum and macrophages, which could contribute to attenuation of atherosclerosis development in these patients.


      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


        • Ziegler D.
        Type 2 diabetes as an inflammatory cardiovascular disorder.
        Curr Mol Med. 2005; 5: 309-322
        • Wattanakit K.
        • Folsom A.R.
        • Selvin E.
        • et al.
        Risk factors for peripheral arterial disease incidence in persons with diabetes: the Atherosclerosis Risk in Communities (ARIC) Study.
        Atherosclerosis. 2005; 180: 389-397
        • Aronson D.
        • Rayfield E.J.
        How hyperglycemia promotes atherosclerosis: molecular mechanisms.
        Cardiovasc Diabetol. 2002; 1: 1
        • Ceriello A.
        • Motz E.
        Is oxidative stress the pathogenic mechanism underlying insulin resistance, diabetes, and cardiovascular disease? The common soil hypothesis revisited.
        Arterioscler Thromb Vasc Biol. 2004; 24: 816-823
        • Martin-Gallan P.
        • Carrascosa A.
        • Gussinye M.
        • Dominguez C.
        Biomarkers of diabetes-associated oxidative stress and antioxidant status in young diabetic patients with or without subclinical complications.
        Free Radic Biol Med. 2003; 34: 1563-1574
        • Whiteside C.L.
        Cellular mechanisms and treatment of diabetes vascular complications converge on reactive oxygen species.
        Curr Hypertens Rep. 2005; 7: 148-154
        • Johansen J.S.
        • Harris A.K.
        • Rychly D.J.
        • Ergul A.
        Oxidative stress and the use of antioxidants in diabetes: linking basic science to clinical practice.
        Cardiovasc Diabetol. 2005; 4: 5
        • Park S.
        • Choi S.B.
        Effects of alpha-tocopherol supplementation and continuous subcutaneous insulin infusion on oxidative stress in Korean patients with type 2 diabetes.
        Am J Clin Nutr. 2002; 75: 728-733
        • Ceriello A.
        • Bortolotti N.
        • Motz E.
        • et al.
        Red wine protects diabetic patients from metal-induced oxidative stress and thrombosis: a pleasant approach to the prevention of cardiovascular disease in diabetes.
        Eur J Clin Invest. 2001; 31: 322-328
        • Rizvi S.I.
        • Zaid M.A.
        • Anis R.
        • Mishra N.
        Protective role of tea catechins against oxidation-induced damage of type 2 diabetic erythrocytes.
        Clin Exp Pharmacol Physiol. 2005; 32: 70-75
        • Gil M.L.
        • Tomas-Barberan F.A.
        • Hess-Pierce B.
        • Holcroft D.M.
        • Kader A.A.
        Antioxidant activity of pomegranate juice and its relationship with phenolics composition and processing.
        J Agric Food Chem. 2000; 10: 4581-4589
      1. Aviram M. Pomegranate juice as a major source for polyphenolic flavonoids and it is most potent antioxidant against LDL oxidation and atherosclerosis. In: Pasquier C, editor. Proceedings of the XI Biennial Meeting of the Society for Free Radical Research International. Paris, France, 16–20 July, 2002. Monduzzi Editore S.P, A-MEDIMOND Inc., pp. 523–28.

        • Aviram M.
        • Dornfeld L.
        • Rosenblat M.
        • et al.
        Pomegranate juice consumption reduces oxidative stress, atherogenic modifications to LDL, and platelet aggregation: studies in humans and in atherosclerotic apolipoprotein E-deficient mice.
        Am J Clin Nutr. 2000; 71: 1062-1076
        • Aviram M.
        • Rosenblat M.
        • Gaitini D.
        • et al.
        Pomegranate juice consumption for 3 years by patients with carotid artery stenosis reduces common carotid intima-media thickness, blood pressure and LDL oxidation.
        Clin Nutr. 2004; 23: 423-433
        • Boyle J.J.
        Macrophage activation in atherosclerosis: pathogenesis and pharmacology of plaque rupture.
        Curr Vasc Pharmacol. 2005; 3: 63-68
        • Naito T.
        • Oikawa S.
        • Kotake H.
        • Hayasaka K.
        • Toyota T.
        Effect of glucose concentration on foam cell formation in THP-1 cells.
        J Atheroscler Thromb. 2001; 8: 55-62
        • Dobrian A.D.
        • Lazar V.
        • Sinescu C.
        • Mincu D.
        • Simionescu M.
        Diabetic state induces lipid loading and altered expression and secretion of lipoprotein lipase in human monocytes-derived macrophages.
        Atherosclerosis. 2000; 153: 191-201
        • Ceolotto G.
        • Gallo A.
        • Miola M.
        • et al.
        Protein kinase C activity is acutely regulated by plasma glucose concentration in human monocytes in vivo.
        Diabetes. 1999; 48: 1316-1322
        • Ling Li
        • Tatsuya S.
        • Genevieve R.
        Glucose enhances human macrophage LOX-1 expression: role for LOX-1 in glucose-induced macrophage foam cell formation.
        Circ Res. 2004; 94: 892-901
        • Fukuhara-Takaki K.
        • Sakai M.
        • Sakamoto Y.
        • Takeya M.
        • Horiuchi S.
        Expression of class A scavenger receptor is enhanced by high glucose in vitro under diabetic conditions in vivo: one mechanism for an increased rate of atherosclerosis in diabetes.
        J Biol Chem. 2005; 280: 3355-3364
      2. Hayek T, Hussein K, Aviram M, et al. Macrophage foam cell formation in streptozotocin-induced diabetic mice: stimulatory effect of glucose. Atherosclerosis, in press.

        • Gan K.N.
        • Smolen A.
        • Eckerson H.W.
        • La DU B.N.
        Purification of human serum paraoxonase/arylesterase. Evidence for one esterase catalyzing both activities.
        Drug Metab Dispos. 1991; 19: 100-106
        • Frei B.
        • Stocker R.
        • Ames B.N.
        Antioxidant defenses and lipid peroxidation in human blood plasma.
        Proc Natl Acad Sci USA. 1988; 85: 9748-9752
        • Buege J.A.
        • Aust S.D.
        Microsomal lipid peroxidation.
        Methods Enzymol. 1978; 52: 302-310
        • El-Saadani M.
        • Esterbauer N.
        • El-Sayed M.
        • Goher M.
        • Nassar A.Y.
        • Jurgens G.
        Spectrophotometric assay for lipid peroxides in serum lipoproteins using commercially available reagent.
        J Lipid Res. 1989; 30: 627-630
        • Hu M.-L.
        • Dillard C.J.
        • Tappel A.I.
        In vivo effects of aurothioglucose and sodium thioglucose on rat tissue sulfhydryl levels and plasma sulfhydryl reactivity.
        Agents Actions. 1988; 25: 132-138
        • LeBel C.P.
        • Ischiropoulos H.
        • Bondy S.C.
        Evaluation of the probe 2′,7′-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress.
        Chem Res Toxicol. 1992; 5: 227-231
        • Bass D.A.
        • Parce J.W.
        • Dechatelet L.R.
        • Szejda P.
        • Seeds M.C.
        • Thomas M.
        Flow cytometric studies of oxidative product formation by neutrophils: a graded response to membrane stimulation.
        J Immunol. 1983; 130: 1910-1917
        • Lowry O.H.
        • Rosebrough N.J.
        • Farr A.L.
        • Randall R.J.
        Protein measurement with the Folin phenol reagent.
        J Biol Chem. 1951; 193: 265-275
        • Tietze F.
        Enzymatic method for quantitative determination of nanogram amounts of total and oxidized glutathione: application to mammalian blood and other tissues.
        Anal Biochem. 1969; 27: 502-522
        • Aviram M.
        Plasma lipoprotein separation by discontinuous density gradient ultracentrifugation in hyperlipoproteinemic patients.
        Biochem Med. 1983; 30: 111-118
        • Aviram M.
        • Rosenblat M.
        Paraoxonases 1, 2 and 3, oxidative stress and macrophage foam cells formation during atherosclerosis development.
        Free Radic Biol Med. 2004; 37: 1304-1316
        • Sies H.
        Glutathione and its role in cellular functions.
        Free Radic Res. 1999; 27: 916-921
        • Fuhrman B.
        • Volkova N.
        • Aviram M.
        Oxidative stress increases the expression of the CD36 scavenger receptor and the cellular uptake of oxidized LDL in macrophage from atherosclerotic mice: protective role of antioxidants and paraoxonase.
        Atherosclerosis. 2002; 161: 307-316
        • Krentz A.J.
        Lipoprotein abnormalities and their consequences for patients with type 2 diabetes.
        Diab Obes Metab. 2003; 5: S19-S27
        • El-Nemr S.E.
        • Ismail I.A.
        • Ragab M.
        Chemical composition of juice and seeds of pomegranate fruit.
        Nahrung. 1991; 34: 601-606
        • Ozdemir G.
        • Ozden M.
        • Maral H.
        • Kuskay S.
        • Cetinalp P.
        • Tarkum I.
        Malondialdehyde, glutathione, glutathione peroxidase and homocysteine levels in type 2 diabetic patients with and without microalbuminuria.
        Am Clin Biochem. 2005; 42: 99-104
        • Knott H.M.
        • Brown B.E.
        • Davies M.J.
        • Dean R.T.
        Glycation and glycooxidation of low-density lipoproteins by glucose and low-molecular mass aldehydes. Formation of modified and oxidized particles.
        Eur J Biochem. 2003; 270: 3572-3582
        • Gowri M.S.
        • Van der Westhuyzen D.R.
        • Brodges S.R.
        • Anderson J.W.
        Decreased protection by HDL from poorly controlled type 2 diabetic subjects against LDL oxidation may be due to the abnormal composition of HDL.
        Arterioscler Thromb Vasc Biol. 1999; 19: 2226-2233
        • Letellier C.
        • Duron M.R.
        • Jouanolle A.M.
        • et al.
        Serum paraoxonase activity and paraoxonase gene polymorphism in type 2 diabetic patients with or without vascular complications.
        Diab Metab. 2002; 28: 297-304
        • Tsuzura S.
        • Ikeda Y.
        • Suchiro T.
        • et al.
        Correlation of plasma oxidized low-density lipoprotein levels to vascular complications and human serum paraoxonase in patients with type 2 diabetes.
        Metabolism. 2004; 53: 297-302
        • Aviram M.
        • Rosenblat M.
        • Billecke S.
        • et al.
        Human serum paraoxonase (PON 1) is inactivated by oxidized low density lipoprotein and preserved by antioxidants.
        Free Radic Biol Med. 1999; 26: 892-904
        • Ferretti G.
        • Bacchetti T.
        • Marchionni C.
        • et al.
        Effect of glycation of high density lipoproteins on their physiochemical properties and on paraoxonase activity.
        Acta Diabetol. 2001; 38: 163-169
        • Rozenberg O.
        • Shih D.M.
        • Aviram M.
        Paraoxonase (1) attenuates macrophage oxidative status: studies in PON1 transfected cells and in PON1 transgenic mice.
        Atherosclerosis. 2005; 181: 9-18
        • Fuhrman B.
        • Volkova N.
        • Aviram M.
        Pomegranate juice inhibits oxidized LDL uptake and cholesterol biosynthesis in macrophages.
        Atherosclerosis. 2005; 16: 570-576
        • Sampson M.J.
        • Davies I.R.
        • Braschi S.
        • Ivory K.
        • Hughes D.A.
        Increased expression of scavenger receptor (CD36) in monocytes from subjects with type 2 diabetes.
        Atherosclerosis. 2003; 167: 120-134
        • Kaplan M.
        • Williams K.J.
        • Mandel H.
        • Aviram M.
        Role of macrophage glycosaminoglycans in the cellular catabolism of oxidized LDL by macrophages.
        Arterioscler Thromb Vasc Biol. 1998; 4: 542-553
        • Sah J.F.
        • Balasubramaniam S.
        • Ecker R.L.
        • Rorke E.A.
        Epigallocatechin-3-gallate inhibits epidermal grpwth factor receptor signaling pathway. Evidence for direct inhibition of ERK1/2 and AKT kinases.
        J Biol Chem. 2004; 279: 12755-12762