Molecular and Cellular Biochemistry

, Volume 339, Issue 1–2, pp 55–61

Carnosine supplementation protects rat brain tissue against ethanol-induced oxidative stress

  • Ummuhani Ozel Turkcu
  • Ayşe Bilgihan
  • Gursel Biberoglu
  • Oznur Mertoglu Caglar
Article

Abstract

Ethanol causes oxidative stress and tissue damage. The aim of this study was to investigate the effect of antioxidant carnosine on the oxidative stress induced by ethanol in the rat brain tissue. Forty male rats were divided equally into four groups as control, carnosine (CAR), ethanol (EtOH), and ethanol plus carnosine (EtOH + CAR). Rats in the control group (n = 10) were injected intraperitoneally (i.p.) with 0.9% saline; EtOH group (n = 10) with 2 g/kg/day ethanol, CAR group (n = 10) received carnosine at a dose of 1 mg/kg/day and EtOH + CAR group (n = 10) received carnosine (orally) and ethanol (i.p.). All animals were sacrificed using ketamine and brain tissues were removed. Malondialdehyde (MDA), protein carbonyl (PCO) and tissue carnosine levels, and superoxide dismutase (SOD) activities were measured. Endogenous CAR levels in the rat brain tissue specimens were significantly increased in the CAR and EtOH groups when compared to the control animals. MDA and PCO levels in the EtOH group were significantly increased as compared to the other groups (P < 0.05). CAR treatment also decreased MDA levels in the CAR group as compared to the control group. Increased SOD activities were obtained in the EtOH + CAR group as compared to the control (P < 0.05). CAR levels in the rat brain were significantly increased in the CAR, EtOH and CAR + EtOH groups when compared to the control animals. These findings indicated that carnosine may appear as a protective agent against ethanol-induced brain damage.

Keywords

Ethanol Carnosine Brain Oxidative stress Protein carbonyls 

References

  1. 1.
    Pohorecky LA, Brick J (1998) Pharmacology of ethanol. Pharmacol Ther 36(2–3):335–427Google Scholar
  2. 2.
    Zimatkin SM, Deitrich RA (1997) Ethanol metabolism in the brain. Addict Biol 2:387–392CrossRefGoogle Scholar
  3. 3.
    Bora PS, Lange LG (1993) Molecular mechanism of ethanol metabolism by human brain to fatty acid ethyl esters. Alcohol Clin Exp Res 17:28–30CrossRefPubMedGoogle Scholar
  4. 4.
    El-Sokkary GH, Reiter RJ, Tan DX, Kim SJ, Cabrera J (1999) Inhibitory effect of melatonin on products of lipid peroxidation resulting from chronic ethanol administration. Alcohol Alcohol 34:842–850PubMedGoogle Scholar
  5. 5.
    Renis M, Calabrese V, Russo A, Calderone A, Barcellona ML, Rizza V (1996) Nuclear DNA strand breaks during ethanol-induced oxidative stress in rat brain. FEBS Lett 390:153–156CrossRefPubMedGoogle Scholar
  6. 6.
    Calabrese V, Renis M, Calderone A, Russo A, Reale S, Barcellona ML, Rizza V (1998) Stress proteins and SH-groups in oxidant-induced cellular injury after chronic ethanol administration in rat. Free Radic Biol Med 24:1159–1167CrossRefPubMedGoogle Scholar
  7. 7.
    Skaper SD, Floreani M, Ceccon M, Facci L, Giusti P (1999) Excitotoxicity, oxidative stress, and the neuroprotective potential of melatonin. Ann NY Acad Sci 890:107–118CrossRefPubMedGoogle Scholar
  8. 8.
    Jennett RB, Sorrell MF, Johnson EL, Tuma DJ (1987) Covalent binding of acetaldehyde to tubulin: evidence for preferential binding to the alpha-chain. Arch Biochem Biophys 256:10–18CrossRefPubMedGoogle Scholar
  9. 9.
    Kenney WC (1982) Acetaldehyde adducts of phospholipids. Alcohol Clin Exp Res 6:412–416CrossRefPubMedGoogle Scholar
  10. 10.
    Rintala J, Jaatinen P, Parkkila S, Sarviharju M, Kiianmaa K, Hervonen A, Niemelä O (2000) Evidence of acetaldehyde-protein adduct formation in rat brain after lifelong consumption of ethanol. Alcohol Alcohol 35(5):458–463PubMedGoogle Scholar
  11. 11.
    Montoliu C, Vallés S, Renau-Piqueras J, Guerri C (1994) Ethanol-induced oxygen radical formation and lipid peroxidation in rat brain: effect of chronic alcohol consumption. J Neurochem 63(5):1855–1862PubMedGoogle Scholar
  12. 12.
    Montoliu C, Sancho-Tello M, Azorin I, Burgal M, Vallés S, Renau-Piqueras J, Guerri C (1995) Ethanol increases cytochrome P4502E1 and induces oxidative stress in astrocytes. J Neurochem 65(6):2561–2570PubMedGoogle Scholar
  13. 13.
    Ekström G, Ingelman-Sundberg M (1989) Rat liver microsomal NADPH-supported oxidase activity and lipid peroxidation dependent on ethanol-inducible cytochrome P-450 (P-450IIE1). Biochem Pharmacol 38(8):1313–1319CrossRefPubMedGoogle Scholar
  14. 14.
    Sun AY, Sun GY (2001) Ethanol and oxidative mechanisms in the brain. J Biomed Sci 8:37–43CrossRefPubMedGoogle Scholar
  15. 15.
    Abuja PM, Albertini R (2001) Methods for monitoring oxidative stress, lipid peroxidation and oxidation resistance of lipoproteins. Clin Chim Acta 306:1–17CrossRefPubMedGoogle Scholar
  16. 16.
    Smith CD, Carney JM, Starke-Reed PE, Oliver CN, Stadtman ER, Floyd RA, Markesbery WR (1991) Excess brain protein oxidation and enzyme dysfunction in normal aging and in Alzheimer disease. Proc Natl Acad Sci USA 88:10540–10543CrossRefPubMedGoogle Scholar
  17. 17.
    Floor E, Wetzel MG (1998) Increased protein oxidation in human substantia nigra pars compacta in comparison with basal ganglia and prefrontal cortex measured with an improved dinitrophenylhydrazine assay. J Neurochem 70(1):268–275PubMedCrossRefGoogle Scholar
  18. 18.
    Freeman BA, Crapo JD (1982) Biology of disease, free radicals and tissue injury. Lab Investig 47:412–426PubMedGoogle Scholar
  19. 19.
    Boldyrev AA, Severin SE (1990) The histidine-containing dipeptides, carnosine and anserine: distribution, properties and biological significance. Adv Enzym Regul 30:175–194CrossRefGoogle Scholar
  20. 20.
    Pavlov AR, Revina AA, Dupin AM, Boldyrev AA, Yaropolov AL (1993) The mechanism of interaction of carnosine with superoxide radicals in water solutions. Biochim Biophys Acta 1157(3):304–312PubMedGoogle Scholar
  21. 21.
    Kohen R, Misgav R, Ginsburg I (1991) The SOD like activity of copper:carnosine, copper:anserine and copper:homocarnosine complexes. Free Radic Res Commun 12–13:179–185CrossRefPubMedGoogle Scholar
  22. 22.
    Gallant S, Kukley M, Stvolinsky S, Bulygina E, Boldyrev A (2000) Effect of carnosine on rats under experimental brain ischemia. Tohoku J Exp Med 191(2):85–99CrossRefPubMedGoogle Scholar
  23. 23.
    Deev LI, Goncharenko EN, Baĭzhumanov AA, Akhalaia MI, Antonova SV, Shestakova SV (1997) Protective effect of carnosine in hyperthermia. Bull Eksp Biol Med 124(7):50–52Google Scholar
  24. 24.
    Münch G, Mayer S, Michaelis J, Hipkiss AR, Riederer P, Müller R, Neumann A, Schinzel R, Cunningham AM (1997) Influence of advanced glycation end-products and AGE-inhibitors on nucleation-dependent polymerization of beta-amyloid peptide. Biochim Biophys Acta 1360(1):17–29PubMedGoogle Scholar
  25. 25.
    Hipkiss AR, Preston JE, Himswoth DT, Worthington VC, Abbot NJ (1997) Protective effects of carnosine against malondialdehyde-induced toxicity towards cultured rat brain endothelial cells. Neurosci Lett 283(3):135–138CrossRefGoogle Scholar
  26. 26.
    Nagasawa T, Yonekura T, Nishizawa N, Kitts DD (2001) In vitro and in vivo inhibition of muscle lipid and protein oxidation by carnosine. Mol Cell Biochem 225(1):29–34CrossRefPubMedGoogle Scholar
  27. 27.
    Brownson C, Hipkiss AR (2000) Carnosine reacts with a glycated protein. Free Radic Biol Med 28:1564–1570CrossRefPubMedGoogle Scholar
  28. 28.
    Hipkiss AR, Brownson C (2000) Carnosine reacts with protein carbonyl groups: another possible role for the anti-ageing peptide? Biogerontology 1:217–223CrossRefPubMedGoogle Scholar
  29. 29.
    Sun Y, Oberley LW, Li Y (1988) A simple method for clinical assay of superoxide dismutase. Clin Chem 34(13):497–500PubMedGoogle Scholar
  30. 30.
    Reznick AZ, Packer L (1994) Oxidative damage to proteins: spectrophotometric method for carbonyl assay. Methods Enzymol 233:357–363CrossRefPubMedGoogle Scholar
  31. 31.
    Dunnett M, Harris RC (1977) High-performance liquid chromatography determination of imidazole dipeptides, histidines, 1-methylhistidine and 3-methylhistidine in equine and camel muscle and individual muscle fibre. J Chromatogr B Biochem Appl 688:47–55CrossRefGoogle Scholar
  32. 32.
    Kasiba E, Flancbaum L, Fitzpatrick JC, Schneidrman J, Fisher H (1988) Simultaneous determination of histidine-containing dipeptides, histamine, methylhistamine and histidine by high-performance liquid chromatography. J Chromatogr 432:315–320CrossRefGoogle Scholar
  33. 33.
    Ceconi C, Cargnoni A, Pasini E, Condorelli E, Curello S, Ferrari R (1991) Evaluation of phospholipid peroxidation as malondialdehyde during myocardial ischemia and reperfusion injury. Am J Physiol 260(4):H:1057–H:1061Google Scholar
  34. 34.
    Cini M, Moretti A (1995) Business Unit Therapeutics Preclinical Research, Pharmacia Farmitalia Carlo Erba Nerviano Italy. Studies on lipid peroxidation and protein oxidation in the aging brain. Neurobiol Aging 16:53–57CrossRefPubMedGoogle Scholar
  35. 35.
    Pilz J, Meineke I, Gleiter CH (2000) Measurement of free and bound malondialdehyde in plasma by high-performance liquid chromatography as the 2,4-dinitrophenylhydrazine derivative. J Chromatogr B Biomed Sci Appl 742(2):315–325CrossRefPubMedGoogle Scholar
  36. 36.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Biochem 193:145–157Google Scholar
  37. 37.
    Pushpakiran G, Mahalakshmi K, Anuradha CV (2004) Taurine restores ethanol-induced depletion of antioxidants and attenuates oxidative stress in rat tissues. Amino Acids 27(1):91–96CrossRefPubMedGoogle Scholar
  38. 38.
    Kurose I, Higuchi H, Kato S, Miura S, Ishii H (1996) Ethanol-induced oxidative stress in the liver. Alcohol Clin Exp Res 20(1):77–85CrossRefGoogle Scholar
  39. 39.
    Nordmann R (1994) Alcohol and antioxidant systems. Alcohol Alcohol 29(5):513–522PubMedGoogle Scholar
  40. 40.
    Kukielka E, Cederbaum AI (1994) DNA strand cleavage as a sensitive assay for the production of hydroxyl radicals by microsomes: role of cytochrome P4502E1 in the increased activity after ethanol treatment. Biochem J 302(3):773–779PubMedGoogle Scholar
  41. 41.
    Wang M, Mcintee EJ, Cheng G (2000) Identification of DNA adducts of acetaldehyde. Chem Res Toxicol 13:1149–1157CrossRefPubMedGoogle Scholar
  42. 42.
    Ostrowska J, Łuczaj W, Kasacka I, Rózański A, Skrzydlewska E (2004) Green tea protects against ethanol-induced lipid peroxidation in rat organs. Alcohol 32(1):25–32CrossRefPubMedGoogle Scholar
  43. 43.
    Nordmann R, Ribie′re C, Rouach H (1992) Implication of free radical mechanisms in ethanol induced cellular injury. Free Radic Biol Med 12:219–240CrossRefPubMedGoogle Scholar
  44. 44.
    Burger RM, Berkowitz AR, Peisach J (1980) Origin of malondialdehyde from DNA degraded by Fe(II) bleomycin. J Biol Chem 255:11832–11838PubMedGoogle Scholar
  45. 45.
    Gutteridge JM (1981) Thiobarbituric acid-reactivity following iron-dependent free-radical damage to amino acids and carbohydrates. FEBS Lett 128:343–347CrossRefPubMedGoogle Scholar
  46. 46.
    Vendemiale G, Altomare E, Grattagliano I, Albano O (1989) Increased plasma levels of glutathione and malondialdehyde after acute ethanol ingestion in humans. J Hepatol 9(3):359–365CrossRefPubMedGoogle Scholar
  47. 47.
    Rouach H, Fataccioli V, Gentil M, French SW, Morimoto M, Nordmann R (1997) Effect of chronic ethanol feeding on lipid peroxidation and protein oxidation in relation to liver pathology. Hepatology 25(2):351–355CrossRefPubMedGoogle Scholar
  48. 48.
    Kanbak G, Arslan OC, Dokumacioglu A, Kartkaya K, Inal ME (2008) Effects of chronic ethanol consumption on brain synaptosomes and protective role of betaine. Neurochem Res 33(3):539–544CrossRefPubMedGoogle Scholar
  49. 49.
    Berlett BS, Stadtman ER (1997) Protein oxidation in aging, disease, and oxidative stress. J Biol Chem 272:20313–20316CrossRefPubMedGoogle Scholar
  50. 50.
    Davies MJ (2005) The oxidative environment and protein damage. Biochim Biophys Acta 1703:93–109PubMedGoogle Scholar
  51. 51.
    Dalle-Donne I, Rossi R, Giustarini D, Milzani A, Colombo R (2003) Protein carbonyl groups as biomarkers of oxidative stress. Clin Chim Acta 329:23–38CrossRefPubMedGoogle Scholar
  52. 52.
    Requena JR, Levine RL, Stadtman ER (2003) Recent advances in the analysis of oxidized proteins. Amino Acids 25(3–4):221–226CrossRefPubMedGoogle Scholar
  53. 53.
    Pirlich M, Piok C, Sandig G, Lochs H, Grune T (2002) Alpha-lipoic acid prevents ethanol-induced protein oxidation in mouse hippocampal HT22 cells. Neurosci Lett 328:93–96CrossRefPubMedGoogle Scholar
  54. 54.
    Ren JC, Banan A, Keshavarzian A, Zhu Q, Lapaglia N, McNulty J, Emanuele NV, Emanuele MA (2005) Exposure to ethanol induces oxidative damage in the pituitary gland. Alcohol 35(2):91–101CrossRefPubMedGoogle Scholar
  55. 55.
    Fataccioli V, Andraud E, Gentil M, French SW, Rouach H (1999) Effects of chronic ethanol administration on rat liver proteasome activities: relationship with oxidative stress. Hepatology 29:14–20CrossRefPubMedGoogle Scholar
  56. 56.
    Kannan M, Wang L, Kang YJ (2004) Myocardial oxidative stress and toxicity induced by acute ethanol exposure in mice. Exp Biol Med (Maywood) 229(6):553–559Google Scholar
  57. 57.
    Yen WJ, Chan LW, C-Pu Lee, Duh P (2002) Inhibition of lipid peroxidation and nonlipid oxidative damage by carnosine. JAOCS 79(4):329–333CrossRefGoogle Scholar
  58. 58.
    Wang H, Marnett LJ, Harris TM, Rizzo CJ (2004) A novel synthesis of malondialdehyde adducts of deoxyguanosine, deoxyadenosine, and deoxycytidine. Chem Res Toxicol 17:144–149CrossRefPubMedGoogle Scholar
  59. 59.
    Hipkiss AR, Worthington VC, Himsworth DT, Herwig W (1998) Protective effects of carnosine against protein modification mediated by malondialdehyde and hypochlorite. Biochim Biophys Acta 1380(1):46–54PubMedGoogle Scholar
  60. 60.
    Hipkiss AR (1998) Carnosine, a protective, anti-ageing peptide? Int J Biochem Cell Biol 30(8):863–868CrossRefPubMedGoogle Scholar
  61. 61.
    McCord JM, Fridovich I (1968) The reduction of cytochrome c by milk xanthine oxidase. J Biol Chem 243:5753–5760PubMedGoogle Scholar
  62. 62.
    Knecht KT, Thurman RG, Mason RP (1993) Role of superoxide and trace transition metals in the production of alpha-hydroxyethyl radical from ethanol by microsomes from alcohol dehydrogenase-deficient deer mice. Arch Biochem Biophys 303(2):339–348CrossRefPubMedGoogle Scholar
  63. 63.
    Badwey JA, Karnovsky ML (1980) Active oxygen species and the functions of phagocytic leukocytes. Annu Rev Biochem 49:695–726CrossRefPubMedGoogle Scholar
  64. 64.
    Fridovich I (1976) Oxygen radicals, hydrogen peroxide and oxygen toxicity. In: Pryor WA (ed) Free radicals in biology. Academic Press, New York, pp 239–277Google Scholar
  65. 65.
    Augustyniak A, Waszkiewicz E, Skrzydlewska E (2005) Preventive action of green tea from changes in the liver antioxidant abilities of different aged rats intoxicated with ethanol. Nutrition 21(9):925–932PubMedGoogle Scholar
  66. 66.
    Son DO, Satsu H, Kiso Y, Shimizu M (2004) Characterization of carnosine uptake and its physiological function in human intestinal epithelial Caco-2 cells. Biofactors 21:395–398CrossRefPubMedGoogle Scholar
  67. 67.
    Gardner LLG, Illingworth KM, Kelleher J, Wood D (1991) Intestinal absorption of the intact peptide carnosine in man, and comparison with intestinal permeability to lactulose. J Physiol 439:411–422PubMedGoogle Scholar
  68. 68.
    Lenney JF, Peppers SC, Kucera-Orallo CM, George RP (1985) Characterization of human tissue carnosinase. Biochem J 228:653–660PubMedGoogle Scholar
  69. 69.
    Tamaki N, Funatsuka A, Fujimoto S, Hama T (1984) The utilization of carnosine in rats fed on a histidine-free diet and its effect on the levels of tissue histidine and carnosine. J. Nutr Sci Vitaminol (Tokyo) 30:541–551Google Scholar
  70. 70.
    Jackson MC, Kucera CM, Lenney JF (1991) Purification and properties of human serum carnosinase. Clin Chim Acta 196(2–3):193–205CrossRefPubMedGoogle Scholar
  71. 71.
    Lenney JF, Peppers SC, Kucera CM, Sjaastad O (1983) Homocarnosinosis: lack of serum carnosinase is the defect probably responsible for elevated brain and CSF homocarnosine. Clin Chim Acta 132(2):157–165CrossRefPubMedGoogle Scholar
  72. 72.
    Duane P, Peters TJ (1988) Serum carnosinase activities in patients with alcoholic chronic skeletal muscle myopathy. Clin Sci (Lond) 75(2):185–190Google Scholar
  73. 73.
    McLoughlin DM, Wassif WS, Morton J, Spargo E, Peters TJ, Russell GF (2000) Metabolic abnormalities associated with skeletal myopathy in severe anorexia nervosa. Nutrition 16(3):192–196CrossRefPubMedGoogle Scholar
  74. 74.
    Abdel-Nabi R, Milakofsky L, Hofford J, Hare TA, Vogel WH (1996) Effect of ethanol on amino acids and related compounds in rat plasma, heart, aorta, bronchus, and pancreas. Alcohol 13(2):171–174CrossRefPubMedGoogle Scholar
  75. 75.
    Stvolinskii SL, Fedorova TN, Yuneva MO, Boldyrev AA (2003) Protective effect of carnosine on Cu, Zn-superoxide dismutase during impaired oxidative metabolism in the brain in vivo. Bull Exp Biol Med 135(2):130–132CrossRefPubMedGoogle Scholar
  76. 76.
    Choi SY, Kwon HY, Kwon OB (1999) Hydrogen peroxide-mediated Cu, Zn-superoxide dismutase fragmentation: protection by carnosine, homocarnosine and anserine. Biochim Biophys Acta 1472(3):651–657PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Ummuhani Ozel Turkcu
    • 1
  • Ayşe Bilgihan
    • 2
  • Gursel Biberoglu
    • 3
  • Oznur Mertoglu Caglar
    • 2
  1. 1.School of Health Sciences, Mugla UniversityMuglaTurkey
  2. 2.Department of Biochemistry, Faculty of MedicineGazi UniversityAnkaraTurkey
  3. 3.Department of Pediatric Metabolism and Nutrition, Faculty of MedicineGazi UniversityAnkaraTurkey

Personalised recommendations