Abstract
Ethanol disorders biological membranes causing perturbations in the bilayer and also by altering the physicochemical properties of membrane lipids. But, chronic alcohol consumption also increases nitric oxide (NO) production. There was no systemic study was done related to alcohol-induced production of NO and consequent formation of peroxynitrite mediated changes in biophysical and biochemical properties, structure, composition, integrity and function of erythrocyte membranes in chronic alcoholics. Hence, keeping all these conditions in mind the present study was undertaken to investigate the role of over produced nitric oxide on red cell membrane physicochemical properties in chronic alcoholics. Human male volunteers aged 44 ± 6 years with similar dietary habits were divided into two groups, namely nonalcoholic controls and chronic alcoholics (~125 g of alcohol at least five times per week for the past 10–12 years). Elevated nitrite and nitrate levels in plasma and lysate, changes in erythrocyte membrane individual phospholipid composition, increased lipid peroxidation, protein carbonyls, cholesterol and phospholipids ratio (C/P ratio) and anisotropic value (γ) with decreased sulfhydryl groups and Na+/K+-ATPase activity in alcoholics was evident from this study. RBC lysate NO was positively correlated with C/P ratio (r = 0.547) and anisotropic (γ) value (r = 0.428), Na+/K+-ATPase activity was negatively correlated with RBC lysate NO (r = −0.372) and anisotropic (γ) value (r = −0.624) in alcoholics. Alcohol-induced overproduction of nitric oxide reacts with superoxide radicals to produce peroxynitrite, which appears to be responsible for changes in erythrocyte membrane lipids and the activity of Na+/K+-ATPase.
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Mansouri A, Demeilliers C, Amsellem S, Pessayre D, Fromenty B (2001) Acute ethanol administration oxidatively damages and depletes mitochondrial DNA in mouse liver, brain, heart and skeletal muscles: protective effects of antioxidants. J Pharmacol Exp Ther 298:731–743
Wu D, Cederbaum AI (2003) Alcohol, oxidative stress, and free radical damage. Alcohol Res Health 27:277–284
Dey A, Cederbaum AI (2006) Alcohol and oxidative liver injury. Hepatology 43:S63–S74
Lindi C, Montorfano G, Marliani P (1998) Rat erythrocyte susceptibility to lipid peroxidation after chronic ethanol intake. Alcohol 16:311–316
Angel C (2009) Lipid peroxidation of membrane phospholipids generates hydroxy-alkenals and oxidized phospholipids active in physiological and/or pathological conditions. Chem Phys Lipids 157:1–11
Taraschi TF, Rubin E (1985) Effects of ethanol on the chemical and structural properties of biologic membranes. Lab Invest 52:120–131
Paramahamsa M, Rameswara Reddy K, Varadacharyulu NC (2004) Changes in composition and properties of erythrocyte membrane in chronic alcoholics. Alcohol Alcoholism 39:110–112
Muriel P, Sandoval G (2000) Nitric oxide and peroxynitrite anion modulate liver plasma membrane fluidity and Na+/K+-ATPase activity. Nitric Oxide 4:333–342
Brzeszczynska J, Gwozdzinski K (2008) Nitric oxide induced oxidative changes in erythrocyte membrane components. Cell Biol Intern 32:114–120
Kleinbongard P, Schulz R, Rassaf T, Lauer T, Dejam A, Jax T, Kumara I, Gharini P, Kabanova S, Ozuyaman B, Schnurch HG, Godecke A, Weber AA, Robenek M, Robenek H, Bloch W, Rosen P, Kelm M (2006) Red blood cells express a functional endothelial nitric oxide synthase. Blood 107:2943–2951
Backman JS, Backman TW, Chen J, Marshall PA, Freeman BA (1990) Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci USA 87:1620–1624
Pacher P, Beckman JS, Liaudet L (2007) Nitric oxide and peroxynitrite in health and disease. Physiol Rev 87:315–424
Devlin TM (1997) Text book of biochemistry with clinical correlations, 4th edn. Wiley-Liss, Inc., New York
Sastry KVH, Moudgal RP, Mohan J, Tyagi JS, Rao GS (2002) Spectrophotometric determination of serum nitrite and nitrate by copper-cadmium alloy. Anal Biochem 306:79–82
Beutler E, Dixon O, Kelly BM (1963) Improved method for the determination of blood glutathione. J Lab Clin Med 61:882–890
Dodge JT, Mitchell C, Hanahan DJ (1963) The preparation and chemical characteristics of hemoglobin free ghosts of human erythrocytes. Arch Biochem Biophys 100:119–130
Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358
Reznick AZ, Packer L (1994) Oxidative damage to proteins: spectroscopic method for carbonyl assay. Methods Enzymol 233:357–363
Sailaja R, Setty OH (2006) Protective effect of phyllanthus fraternus against allyl alcohol-induced oxidative stress in liver mitochondria. J Ethnopharmacol 105:201–209
Ismail B, Edelman IS (1985) Assay of Na+, K+ ATPase. Biochem Pharmacol 34:2685–2689
Folch J, Ascoli I, Lees M, Meath JA, Le-Baron FN (1951) Preparation of lipid extracts from brain tissue. J Biol Chem 191:833–841
Zlatkis A, Zak B, Boyle AJ (1953) A new method for the direct determination of serum cholesterol. J Lab Clin Med 4:486–492
Connerty HV, Briggs AR, Eaton EH (1961) Determination of serum phospholipids, lipid phosphorus. In: Varley H (ed) Practical, clinical biochemistry, 4th edn. CBS publishers, India, pp 319–320
Kavitha G, Damodara Reddy V, Paramahamsa M, Akthar PM, Varadacharyulu NC (2008) Role of nitric oxide in alcohol-induced changes in lipid profile of moderate and heavy alcoholics. Alcohol 42:47–53
Choi JH, Yu BP (1990) Unsuitability of TBA test as a lipid peroxidation marker due to prostaglandin synthesis in the aging kidney. Age 13:61–64
Lowry OH, Rosebrough NJ, Farr AL, Randall R (1951) Protein measurement with the Folin-phenol reagent. J Biol Chem 193:263–275
Foreman DT (1988) The effect of ethanol and its metabolites on carbohydrate, protein and lipid metabolism. Ann Clin Lab Sci 18:181–189
Lieber CS (2000) Alcohol and the liver: metabolism of alcohol and its role in hepatic and extra hepatic diseases. Mt Sinai J Med 67:84–94
Albano E (2006) Alcohol, oxidative stress and free radical damage. Proc Nutr Soc 65:278–290
Zima T, Fialova L, Mestek O, Janebova M, Crkovska J, Malbohan I, Stipek S, Mikullkova L, Poppv P (2001) Oxidative stress, metabolism of ethanol and alcohol-related diseases. J Bio Med Sci 8:59–70
Guajardo M, Terrasa AM, Catala A (2006) Lipid-protein modifications during ascorbate-Fe2+ peroxidation of photoreceptor membranes. Melatonin effect. J Pineal Res 41:201–210
Huang M, Chen C, Peng F, Tang S, Chen C (2009) Alterations in oxidative stress status during early alcohol withdrawal in alcoholic patients. J Formos Med Assoc 108:560–569
Chin JH, Parsons LM, Goldstein DB (1978) Increased cholesterol content of erythrocyte and brain membranes in ethanol-tolerant mice. Biochim Biophys Act 513:358–363
Kanbak G, Akyuz F, Inal M (2001) Preventive effect of betaine on ethanol-induced membrane lipid composition and membrane ATPases. Arch Toxicol 75:59–61
Alling C, Gustavsson L, Mansson JE, Benthin G, Anggard E (1984) Phosphotidylethanol formation in rat organs after ethanol treatment. Biochim Biophys Acta 793:119–122
Mueller GC, Fleming MF, Le Mahieu MA, Lybrand GS, Barry KJ (1988) Synthesis of phosphotidylethanol—a potential marker for adult males at risk for alcoholism. Proc Nat Acad Sci 85:9778–9782
Sale FO, Lindi C, Palestini P, Masserini M (1989) Role of phosphotidyl ethanol is membranes. Effect on membrane fluidity, tolerance to ethanol, and activity of membrane-bound enzymes. Biochemistry 21:1477–1482
Tsuda K, Shimamoto YK, Kimura K, Nishio I (2003) Nitric oxide is a determinant of membrane fluidity of erythrocytes in postmenopausal woman: an electron paramagnetic resonance investigation. Am J Hyper 16:244–248
Hrelia S, Lercker G, Biagi PL, Bordoxi A, Stefanini F, Lunarelli P, Rossi CA (1986) Effect of ethanol intake on human erythrocyte membrane fluidity and lipocomposition. Biochem Intern 12:741–750
Stibler H, Beauge F, Leguicher A, Borg S (1991) Biophysical and biochemical alterations in erythrocyte membranes from chronic alcoholics. Scand J Clin Lab Invest 51:309–319
Johnson JH, Crider BP (1989) Increases in Na+, K+ ATPase activity of erythrocytes and skeletal muscle after chronic ethanol consumption: evidence for reduced efficiency of the enzyme. Proc Natl Acad Sci USA 86:7857–7860
Ishizaki M, Teraoka K, Truritani I, Honda R, Ishida M, Yamada Y (1994) Erythrocyte Na+, K+-ATPase and membrane and serum lipid profiles: as related to alcohol, body mass index and blood pressure. Clin Exp Hyperten 16:741–759
Sutherland E, Dixon BS, Leffert HL, Skally H, Zaccaro L, Simon FR (1988) Biochemical localization of hepatic surface membrane Na+, K+-ATPase activity depends on membrane lipid fluidity. Proc Natl Acad Sci USA 85:8673–8677
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This study was supported in part by the University Grants Commission (Grant No. F-3-11/97), New Delhi, India.
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Maturu, P., Vaddi, D.R., Pannuru, P. et al. Alterations in erythrocyte membrane fluidity and Na+/K+-ATPase activity in chronic alcoholics. Mol Cell Biochem 339, 35–42 (2010). https://doi.org/10.1007/s11010-009-0367-z
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DOI: https://doi.org/10.1007/s11010-009-0367-z