Advertisement

Anti-oxidant and anti-atherogenic properties of liposomal glutathione: Studies in vitro, and in the atherosclerotic apolipoprotein E-deficient mice

      Abstract

      Liposomal glutathione, but not the control liposomes (with no glutathione), dose-dependently inhibited copper ion-induced low density lipoprotein (LDL) and HDL oxidation. As peroxidase activity was found to be present in both LDL and HDL, it has contributed to the anti-oxidative effects of liposomal glutathione. In-vitro, no significant effect of liposomal glutathione on J774 A.1 macrophage cell-line oxidative stress and on cellular cholesterol metabolism was observed. In contrast, in the atherosclerotic apolipoprotein E-deficient (E0) mice, consumption of liposomal glutathione (12.5 or 50 mg/kg/day, for 2 months), but not control liposomes, resulted in a significant reduction in the serum susceptibility to AAPH-induced oxidation by 33%. Liposomal glutathione (50 mg/kg/day) consumption also resulted in an increment (by 12%) in the mice peritoneal macrophages (MPM) glutathione content, paralleled by a significant reduction in total cellular lipid peroxides content (by 40%), compared to placebo-treated mice MPM. MPM paraoxonase 2 activity was significantly increased by 27% and by 121%, after liposomal glutathione consumption (12.5 or 50 mg/kg/day, respectively). Analyses of cellular cholesterol fluxes revealed that, liposomal glutathione (12.5 mg/kg/day) consumption, decreased the extent of oxidized-LDL (Ox-LDL) uptake by 17% and the cellular cholesterol biosynthesis rate, by 34%, and stimulated HDL-induced macrophage cholesterol efflux, by 19%. Most important, a significant reduction in macrophage cholesterol mass (by 24%), and in the atherosclerotic lesion area (by 30%) was noted.
      We thus conclude that liposomal glutathione possesses anti-oxidative and anti-atherogenic properties towards lipoproteins and macrophages, leading to attenuation of atherosclerosis development.

      Keywords

      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:

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

      References

        • Lusis A.J.
        Atherosclerosis.
        Nature. 2000; 404: 233-241
        • Aviram M.
        • Rosenblat M.
        Oxidative stress in cardiovascular disease: role of oxidized lipoproteins in macrophage foam cell formation and atherosclerosis.
        in: Redox-Genome Interactions in Health and Disease. Dekker, New York2003: 57-590 (Chapter 25)
        • Schulze P.C.
        • Lee R.T.
        Oxidative stress and atherosclerosis.
        Curr Atheroscler Rep. 2005; 7: 242-248
        • Hayek T.
        • Oikinine J.
        • Brook J.G.
        • Aviram M.
        Increased plasma and lipoprotein lipid peroxidation in apoE deficient mice.
        Biochem Biophys Res Commun. 1994; 201: 1567-1574
        • Maor I.
        • Kaplan M.
        • Hayek T.
        • Vaya Y.
        • Hoffman A.
        • Aviram M.
        Oxidized monocytes-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
        • Rosenblat M.
        • Hayek T.
        • Aviram M.
        Anti-oxidative effects of pomegranate juice (PJ) consumption by diabetic patients on serum and on macrophages.
        Atherosclerosis. 2006; 187: 363-371
        • Toshima S.
        • Hasegawa A.
        • Kurabayashi M.
        • et al.
        Circulating oxidized low density lipoprotein levels: a biochemical risk marker for coronary heart disease.
        Arterioscler Thromb Vasc Biol. 2000; 20: 2243-2247
        • Fuhrman B.
        • Volkova N.
        • Aviram M.
        Oxidative stress increases the expression of the CD36 scavenger receptors and the cellular uptake of oxidized LDL in macrophages from atherosclerotic mice: protective role of antioxidants and paraoxonase.
        Atherosclerosis. 2002; 161: 307-316
        • Sies H.
        Glutathione and its role in cellular functions.
        Free Radic Biol Med. 1999; 27: 916-921
        • Schafer F.Q.
        • Buettner G.R.
        Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple.
        Free Radic Biol Med. 2001; 30: 1191-1212
        • Ashfaq S.
        • Abramson J.L.
        • Jones D.P.
        • et al.
        The relationship between plasma levels of oxidized and reduced thiols and early atherosclerosis in healthy adults.
        J Am Coll Cardiol. 2006; 47: 1005-1011
        • Biswas S.K.
        • Newby D.E.
        • Rahman I.
        • Megson I.L.
        Decreased glutathione synthesis precedes oxidative stress and atherogenesis in Apo-E (−/−) mice.
        Biochem Biophys Res Commun. 2005; 338: 1368-1373
        • Rosenblat M.
        • Coelman R.
        • Aviram M.
        Increased macrophage glutathione content reduces cell-mediated oxidation of LDL and atherosclerosis in apolipoprotein E-deficient mice.
        Atherosclerosis. 2002; 163: 17-28
        • Sagrista M.L.
        • Garcia A.E.
        • Africa De Madariage M.
        • Mora M.
        Antioxidant and pro-oxidant effect of the thiolic compound N-acetyl-l-cysteine and glutathione against free radical-induced lipid peroxidation.
        Free Radic Res. 2002; 36: 329-340
        • Rosenblat M.
        • Aviram M.
        Macrophage glutathione content and glutathione peroxidase activity are inversely related to cell-mediated oxidation of LDL; in vitro and in vivo studies.
        Free Radic Biol Med. 1998; 24: 305-371
        • Fraternale A.
        • Paoletti M.F.
        • Casabianca A.
        • et al.
        Antiviral and immunomodulatory properties of new pro-glutathione (GSH) molecules.
        Curr Med Chem. 2006; 13: 1749-1755
        • Yanmei W.
        • Qiao M.
        • Mieyal J.J.
        • Asmis L.M.
        • Asmis R.
        Molecular mechanism of glutathione-mediated protection from oxidized low-density lipoprotein-induced cell injury in human macrophages: role of glutathione reductase and glutaredoxin.
        Free Radic Biol Med. 2006; 41: 775-785
        • Rajasekaran N.S.
        • Sathyanarayanan S.
        • Devaraj N.S.
        • Devaraj H.
        Chronic depletion of glutathione (GSH) and minimal modification of LDL in vivo: its prevention by glutathione mono ester (GME) therapy.
        Biochim Biophys Acta. 2005; 1741: 103-112
        • Cooke R.W.
        • Drury J.A.
        Reduction of oxidative stress marker in lung fluid of preterm infants after administration of intra-tracheal liposomal glutathione.
        Biol Neonate. 2005; 87: 178-180
        • Liddell J.R.
        • Dringen R.
        • Crack P.J.
        • Robinson S.R.
        Glutathione peroxidase 1 and a high cellular glutathione concentration are essential for effective organic hydroperoxide detoxification in astrocytes.
        Glia. 2006; 54: 873-879
        • Lin C.C.
        • Yin M.C.
        • Hsu C.C.
        • Lin M.P.
        Effect of five cysteine-containing compounds on three lipogenic enzymes in BALB/cA mice consuming high saturated fat diet.
        Lipids. 2004; 39: 843-848
        • Hassan A.S.
        • Bunick D.
        • St Denis S.H.
        • Lund L.A.
        Glutathione and bile acid synthesis. II. Effect of hepatic glutathione content on the activity and mRNA levels of cholesterol 7 alpha-hydroxylase in the rat.
        Biochem Pharmacol. 1993; 46: 555-556
        • Mathur S.N.
        • Born E.
        • Murthy S.
        • Field F.J.
        Phosphatidylcholine increases the secretion of triacylglycerol-rich lipoproteins by CaCo-2 cells.
        Biochem J. 1996; 314: 569-575
        • Hsu C.C.
        • Huang C.N.
        • Hung Y.C.
        • Yin M.C.
        Five cysteine-containing compounds have antioxidative activity in Balb/cA mice.
        J Nutr. 2004; 134: 149-152
        • Biswas S.K.
        • Newby D.E.
        • Rahman I.
        • Megson I.L.
        Depressed glutathione synthesis precedes oxidative stress and atherogenesis in Apo-E(−/−) mice.
        Biochem Biophys Res Commun. 2005; 338: 1368-1373
        • Rozenberg O.
        • Aviram M.
        S-Glutathionylation regulates HDL-associated paraoxonase 1 (PON1) activity.
        Biochem Biophys Res Commun. 2006; 352: 492-498
        • Klatt P.
        • Lamas S.
        Regulation of protein functions by S-glutathiolation in response to oxidative and nitrosative stress.
        Eur J Biochem. 2000; 267: 4928-4944
        • Ghezzi P.
        • Bonetto V.
        • Fratelli M.
        Thiol-disulfide balance: from the concept of oxidative stress to that of redox regulation.
        Antioxid Redox Signal. 2005; 27: 964-972
        • Schinina M.E.
        • Carlini P.
        • Polticelli F.
        • Zappacosta F.
        • Bossa F.
        • Calabrese L.
        Amino acid sequence of chicken Cu, Zn-containing superoxide dismutase and identification of glutathionyl adducts at exposed cysteine residues.
        Eur J Biochem. 1996; 237: 433-439
        • Ng C.J.
        • Wadleigh D.J.
        • Gangopadhyay A.
        • et al.
        Paraoxonase 2 is a ubiquitously expressed protein with antioxidant properties and is capable of preventing cell-mediated oxidative modification of low density lipoprotein.
        J Biol Chem. 2001; 276: 44444-44449
        • Napolitano M.
        • Rivabene R.
        • Avella M.
        • Botham K.M.
        • Bravo E.
        The internal redox balance of the cells influences the metabolism of lipids of dietary origin by J774 A.1 macrophages: implications for foam cell formation.
        J Vasc Res. 2001; 38: 350-360
        • Bravo E.
        • Napolitano M.
        • Rivabene R.
        Role of pre-existing redox profile of human macrophages on lipid synthesis and cholesterol ester cycle in presence of native, acetylated and oxidized low density lipoprotein.
        J Steroid Biochem Mol Biol. 2001; 77: 73-78
        • Cappel R.E.
        • Gilbert H.F.
        Thiol/disulfide exchange between 3-hydroxy-3-methylglutaryl-CoA reductase and glutathione. A thermodynamically facile dithiol oxidation.
        J Biol Chem. 1988; 263: 12204-12212
        • Schmitz G.
        • Drobnik W.
        ATP-binding cassette transporters in macrophages: promising drug target for treatment of cardiovascular disease.
        Curr Opin Invest Drugs. 2002; 3: 853-888
        • Marcil V.
        • Devin E.
        • Sane A.T.
        • Tremblay A.
        • Levy E.
        Oxidative stress influences cholesterol efflux in THP-1 macrophages: role of ATP-binding cassette A1 and nuclear factors.
        Cardiovasc Res. 2006; 72: 473-482