Molecular and Cellular Biochemistry

, Volume 375, Issue 1–2, pp 39–47 | Cite as

Alcohol-induced oxidative/nitrosative stress alters brain mitochondrial membrane properties

  • Vaddi Damodara Reddy
  • Pannuru Padmavathi
  • Godugu Kavitha
  • Bulle Saradamma
  • Nallanchakravarthula Varadacharyulu


Chronic alcohol consumption causes numerous biochemical and biophysical changes in the central nervous system, in which mitochondria is the primary organelle affected. In the present study, we hypothesized that alcohol alters the mitochondrial membrane properties and leads to mitochondrial dysfunction via mitochondrial reactive oxygen species (mROS) and reactive nitrogen species (RNS). Alcohol-induced hypoxia further enhances these effects. Administration of alcohol to rats significantly increased the mitochondrial lipid peroxidation and protein oxidation with decreased SOD2 mRNA and protein expression was decreased, while nitric oxide (NO) levels and expression of iNOS and nNOS in brain cortex were increased. In addition, alcohol augmented HIF-1α mRNA and protein expression in the brain cortex. Results from this study showed that alcohol administration to rats decreased mitochondrial complex I, III, IV activities, Na+/K+-ATPase activity and cardiolipin content with increased anisotropic value. Cardiolipin regulates numerous enzyme activities, especially those related to oxidative phosphorylation and coupled respiration. In the present study, decreased cardiolipin could be ascribed to ROS/RNS-induced damage. In conclusion, alcohol-induced ROS/RNS is responsible for the altered mitochondrial membrane properties, and alcohol-induced hypoxia further enhance these alterations, which ultimately leads to mitochondrial dysfunction.


Alcohol Cardiolipin Fluidity Hypoxia Mitochondria Nitric oxide 



The authors thank the Director and staff of the Center for Cellular and Molecular Biology, Hyderabad, India, for providing facilities to carry out fluidity studies. This study was supported in part by the University Grants Commission (Grant No. F-3-11/97), New Delhi, India.

Conflict of interest

The authors declare that there are no conflicts of interest.


  1. 1.
    Harper C, Matsumoto I (2005) Ethanol and brain damage. Curr Opin Pharmacol 5:73–78PubMedCrossRefGoogle Scholar
  2. 2.
    Lin MT, Beal MF (2006) Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. Nature 443:787–795PubMedCrossRefGoogle Scholar
  3. 3.
    Almansa I, Fernandez A, García-Ruiz C, Muriach M, Barcia JM, Miranda M, Fernández-Checa JC, Romero FJ (2009) Brain mitochondrial alterations after chronic alcohol consumption. J Physiol Biochem 65:305–312PubMedCrossRefGoogle Scholar
  4. 4.
    Reddy VD, Padmavathi P, Kavitha G, Gopi S, Varadacharyulu N (2011) Emblica officinalis ameliorates alcohol-induced brain mitochondrial dysfunction in rats. J Med Food 14:62–68PubMedCrossRefGoogle Scholar
  5. 5.
    Bailey SM (2003) A review of the role of reactive oxygen and nitrogen species in alcohol-induced mitochondrial dysfunction. Free Radic Res. 37:585–596PubMedCrossRefGoogle Scholar
  6. 6.
    Venkatraman A, Shiva S, Wigley A, Ulasova E, Shhieng D, Bailey SM, Darley-Usmar VM (2004) The role of iNOS in alcohol-dependent hepatotoxicity and mitochondrial dysfunction in mice. Hepatol 40:565–573CrossRefGoogle Scholar
  7. 7.
    Zelickson BR, Benavides GA, Johnson MS, Chacko BK, Venkatraman A, Landar A, Betancourt AM, Bailey SM, Darley-Usmar VM (2011) Nitric oxide and hypoxia exacerbate alcohol-induced mitochondrial dysfunction in hepatocytes. Biochim Biophys Acta 1807:1573–1582PubMedCrossRefGoogle Scholar
  8. 8.
    Deng X, Deitrich RA (2007) Ethanol metabolism and effects: nitric oxide and its interaction. Curr Clin Pharmacol 2:145–153PubMedCrossRefGoogle Scholar
  9. 9.
    Haorah J, Ramirez SH, Floreani N, Gorantla S, Morsey B, Persidsky Y (2008) Mechanism of alcohol-induced oxidative stress and neuronal injury. Free Radic Biol Med 45:1542–1550PubMedCrossRefGoogle Scholar
  10. 10.
    Young TA, Bailey SM, Van Horn CG, Cunningham CC (2006) Chronic ethanol consumption decreases mitochondrial and glycolytic production of ATP in liver. Alcohol 41:254–260Google Scholar
  11. 11.
    Wang SM, Wu R (2009) The double danger of ethanol and hypoxia: their effects on a hepatoma cell line. Int J Clin Exp Pathol 2:182–189PubMedGoogle Scholar
  12. 12.
    Semenza GL (2012) Hypoxia-inducible factors in physiology and medicine. Cell 148:399–408PubMedCrossRefGoogle Scholar
  13. 13.
    Taylor CT, Moncada S (2010) Nitric oxide, cytochrome c oxidase, and the cellular response to hypoxia. Arterioscler Thromb Vasc Biol 30:643–647PubMedCrossRefGoogle Scholar
  14. 14.
    Ball KA, Nelson AW, Foster DG, Poyton RO (2012) Nitric oxide produced by cytochrome c oxidase helps stabilize HIF-1α in hypoxic mammalian cells. Biochem Biophys Res Commun 420:727–732PubMedCrossRefGoogle Scholar
  15. 15.
    Poyton RO, Ball KA, Castello PR (2009) Mitochondrial generation of free radicals and hypoxic signaling. Trends Endocrinol Metab 20:332–340PubMedCrossRefGoogle Scholar
  16. 16.
    Reddy VD, Padmavathi P, Paramahamsa M, Varadacharyulu N (2009) Modulatory role of Emblica officinalis against alcohol induced biochemical and biophysical changes in rat erythrocyte membranes. Food Chem Toxicol 47:1958–1963PubMedCrossRefGoogle Scholar
  17. 17.
    Paradies G, Petrosillo G, Paradies V, Ruggiero FM (2009) Role of cardiolipin peroxidation and Ca2+ in mitochondrial dysfunction and disease. Cell Calcium 45:643–650PubMedCrossRefGoogle Scholar
  18. 18.
    Pfeiffer K, Gohil V, Stuart RA, Hunte C, Brandt U, Greenberg ML, Schägger H (2003) Cardiolipin stabilizes respiratory chain supercomplexes. J Biol Chem 278:52873–52880PubMedCrossRefGoogle Scholar
  19. 19.
    Chicco AJ, Sparagna GC (2007) Role of cardiolipin alterations in mitochondrial dysfunction and disease. Am J Physiol Cell Physiol 292:C33–C44PubMedCrossRefGoogle Scholar
  20. 20.
    Muriel P, Sandoval G (2000) Nitric oxide and peroxynitrite anion modulate liver plasma membrane fluidity and Na+/K+-ATPase activity. Nitric Oxide 4:333–342PubMedCrossRefGoogle Scholar
  21. 21.
    Comellas AP, Dada LA, Lecuona E, Pesce LM, Chandel NS, Quesada N, Budinger GR, Strous GJ, Ciechanover A, Sznajder JI (2006) Hypoxia-mediated degradation of Na, K-ATPase via mitochondrial reactive oxygen species and the ubiquitin-conjugating system. Circ Res 98:1314–1322PubMedCrossRefGoogle Scholar
  22. 22.
    Brustovetsky W, Dubinsky JM (2000) Dual responses of CNS mitochondria to elevated calcium. J Neurosci 20:103–113PubMedGoogle Scholar
  23. 23.
    Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358PubMedCrossRefGoogle Scholar
  24. 24.
    Reznick AZ, Packer L (1994) Oxidative damage to proteins: spectroscopic method for carbonyl assay. Methods Enzymol 233:357–363PubMedCrossRefGoogle Scholar
  25. 25.
    Sastry KHV, Moudgal RP, Mohan J, Tyagi JS, Rao GS (2002) Spectrophotometric determination of serum nitrite and nitrate by copper–cadmium alloy. Anal Biochem 306:79–82PubMedCrossRefGoogle Scholar
  26. 26.
    King TE, Robert HL (1967) Preparations and properties of soluble NADH dehydrogenases from cardiac muscle. In: Estabrook RW, Pullman ME (eds) Methods in enzymology, vol 10. Academic Press Inc., New York, pp 275–294Google Scholar
  27. 27.
    King TE (1967) Preparation of succinate dehydrogenase and reconstitution of succinate oxidase. In: Estabrook RW, Pullman ME (eds) Methods enzymol, vol 10. Academic Press Inc., New York, pp 322–331Google Scholar
  28. 28.
    Krahenbuhl S, Talos C, Wiesmann U, Hoppel CL (1994) Development and evaluation of a spectrophotometric assay for complex III in isolated mitochondria, tissues and fibroblasts from rats and humans. Clin Chim Acta 230:177–187PubMedCrossRefGoogle Scholar
  29. 29.
    Cooperstein SJ, Lazarow A (1951) A microspectrophotometric method for the determination of cytochrome oxidase. J Biol Chem 189:665–670PubMedGoogle Scholar
  30. 30.
    Ismail B, Edelman IS (1985) Assay of Na+, K+-ATPase. Biochem Pharmacol 34:2685–2689CrossRefGoogle Scholar
  31. 31.
    Fiske CH, Subbarow Y (1925) The colorimetric determination of inorganic phosphorus. J Biol Chem 66:375–404Google Scholar
  32. 32.
    Folch J, Ascoli I, Lees M, Meath JA, Le Baron FN (1951) Preparation of lipid extracts from brain tissue. J Biol Chem 191:833–841PubMedGoogle Scholar
  33. 33.
    Padmavathi P, Reddy VD, Kavitha G, Paramahamsa M, Varadacharyulu N (2010) Chronic cigarette smoking alters erythrocyte membrane lipid composition and properties in male human volunteers. Nitric Oxide 23:181–186PubMedCrossRefGoogle Scholar
  34. 34.
    Lowry OH, Rosebrough NJ, Farr AL, Randall R (1951) Protein measurement with the Folin-phenol reagent. J Biol Chem 193:263–275Google Scholar
  35. 35.
    Yuan G, Khan SA, Luo W, Nanduri J, Semenza GL, Prabhakar NR (2011) Hypoxia-inducible factor 1 mediates increased expression of NADPH oxidase-2 in response to intermittent hypoxia. J Cell Physiol 226:2925–2933PubMedCrossRefGoogle Scholar
  36. 36.
    Omodeo-Sale F, Gramigna D, Campaniello R (1997) Lipid peroxidation and antioxidant systems in rat brain: effect of chronic alcohol consumption. Neurochem Res 22:577–582PubMedCrossRefGoogle Scholar
  37. 37.
    Das SK, Hiran KR, Mukherjee S, Vasudevan DM (2007) Oxidative stress is the primary event: effects of ethanol consumption in brain. Indian J Clin Biochem 22:99–104PubMedCrossRefGoogle Scholar
  38. 38.
    Crews FT, Nixon K (2009) Mechanisms of neurodegeneration and regeneration in alcoholism. Alcohol 44:115–127Google Scholar
  39. 39.
    Rump TJ, Abdul Muneer PM, Szlachetka AM, Lamb A, Haorei C, Alikunju S, Xiong H, Keblesh J, Liu J, Zimmerman MC, Jones J, Donohue TM Jr, Persidsky Y, Haorah J (2010) Acetyl-l-carnitine protects neuronal function from alcohol-induced oxidative damage in the brain. Free Radic Biol Med 49:1494–1504PubMedCrossRefGoogle Scholar
  40. 40.
    Catala A (2009) Lipid peroxidation of membrane phospholipids generates hydroxy-alkenals and oxidized phospholipids active in physiological and/or pathological conditions. Chem Phys Lipids 157:1–11PubMedCrossRefGoogle Scholar
  41. 41.
    Davis RL, Syapin PJ (2005) Interactions of alcohol and nitric-oxide synthase in the brain. Brain Res Rev 49:494–504PubMedCrossRefGoogle Scholar
  42. 42.
    Olson N, van der Vliet A (2011) Interactions between nitric oxide and hypoxia-inducible factor signaling pathways in inflammatory disease. Nitric Oxide 25:125–137PubMedCrossRefGoogle Scholar
  43. 43.
    Mantena SK, King AL, Andringa KK, Landar A, Darley-Usmar V, Bailey SM (2007) Novel interactions of mitochondria and reactive oxygen/nitrogen species in alcohol mediated liver disease. World J Gastroenterol 13:4967–4973PubMedGoogle Scholar
  44. 44.
    Nath B, Szabo G (2012) Hypoxia and hypoxia inducible factors: diverse roles in liver diseases. Hepatology 55:622–633PubMedCrossRefGoogle Scholar
  45. 45.
    Mansfield KD, Guzy RD, Pan Y, Young RM, Cash TP, Schumacker PT, Simon MC (2005) Mitochondrial dysfunction resulting from loss of cytochrome c impairs cellular oxygen sensing and hypoxic HIF-alpha activation. Cell Metab 1:393–399PubMedCrossRefGoogle Scholar
  46. 46.
    Padmavathi P, Reddy VD, Maturu P, Varadacharyulu N (2010) Smoking-induced alterations in platelet membrane fluidity and Na(+)/K(+)-ATPase activity in chronic cigarette smokers. J Atheroscler Thromb 17:619–627PubMedCrossRefGoogle Scholar
  47. 47.
    Wiswedel I, Gardemann A, Storch A, Peter D, Schild L (2010) Degradation of phospholipids by oxidative stress-exceptional significance of cardiolipin. Free Radic Res 44:135–145PubMedCrossRefGoogle Scholar
  48. 48.
    Petrosillo G, Portincasa P, Grattagliano I, Casanova G, Matera M, Ruggiero FM, Ferri D, Paradies G (2007) Mitochondrial dysfunction in rat with nonalcoholic fatty liver Involvement of complex I, reactive oxygen species and cardiolipin. Biochim Biophys Acta 1767:1260–1267PubMedCrossRefGoogle Scholar
  49. 49.
    Chandel NS, McClintock DS, Feliciano CE, Wood TM, Melendez JA, Rodriguez AM, Schumacker PT (2000) Reactive oxygen species generated at mitochondrial complex III stabilize hypoxia-inducible factor-1alpha during hypoxia: a mechanism of O2 sensing. J Biol Chem 275:25130–25138PubMedCrossRefGoogle Scholar
  50. 50.
    Castello PR, David PS, McClure T, Crook Z, Poyton RO (2006) Mitochondrial cytochrome oxidase produces nitric oxide under hypoxic conditions: implications for oxygen sensing and hypoxic signaling in eukaryotes. Cell Metab 3:277–287PubMedCrossRefGoogle Scholar
  51. 51.
    Chong PL, Fortes PA, Jameson DM (1985) Mechanisms of inhibition of (Na, K)-ATPase by hydrostatic pressure studied with fluorescent probes. J Biol Chem 260:14484–14490PubMedGoogle Scholar
  52. 52.
    Sutherland E, Dixon BS, Leffert HL, Skally H, Zaccaro L, Simon FR (1998) Biochemical localization of hepatic surface membrane Na+, K+-ATPase activity depends on membrane lipid fluidity. Proc Nat Acad Sci 85:8673–8677CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Vaddi Damodara Reddy
    • 1
    • 2
  • Pannuru Padmavathi
    • 3
  • Godugu Kavitha
    • 1
  • Bulle Saradamma
    • 1
  • Nallanchakravarthula Varadacharyulu
    • 1
  1. 1.Department of BiochemistrySri Krishnadevaraya UniversityAnantapurIndia
  2. 2.Department of MedicinePritzker School of Medicine, University of ChicagoChicagoUSA
  3. 3.Section of Hematology/Oncology, Department of MedicineUniversity of IllinoisChicagoUSA

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