Comparison of Ethanol and Acetaldehyde Toxicity in Rat Astrocytes in Primary Culture

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Comparison of Ethanol and Acetaldehyde Toxicity in Rat Astrocytes in Primary Culture

This study compared the effects of toxicity of ethanol and its first metabolite acetaldehyde in rat astrocytes through cell viability and cell proliferation. The cells were treated with different concentrations of ethanol in the presence or absence of a catalase inhibitor 2-amino-1,2,4 triazole (AMT) or with different concentrations of acetaldehyde. Cell viability was assessed using the trypan blue test. Cell proliferation was assessed after 24 hours and after seven days of exposure to either ethanol or acetaldehyde.

We showed that both ethanol and acetaldehyde decreased cell viability in a dose-dependent manner. In proliferation studies, after seven days of exposure to either ethanol or acetaldehyde, we observed a significant dose-dependent decrease in cell number. The protein content study showed biphasic dose-response curves, after 24 hours and seven days of exposure to either ethanol or acetaldehyde. Co-incubation in the presence of AMT significantly reduced the inhibitory effect of ethanol on cell proliferation.

We concluded that long-term exposure of astrocytes to ethanol is more toxic than acute exposure. Acetaldehyde is a much more potent toxin than ethanol, and at least a part of ethanol toxicity is due to ethanol's first metabolite acetaldehyde.

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  • Singh NP Lai H Khan A. Ethanol-induced single-strand DNA breaks in rat brain cells. Mutat Res 1995;345:191-6.

  • Barret L Soubeyran A Usson Y Eysseric H Saxod R. Characterization of the morphological variations of astrocytes in culture following ethanol exposure. Neurotoxicology 1996;17:497-507.

  • Lamarche F Gonthier B Signorini N Eysseric H Barret L. Acute exposure of cultured neurones to ethanol results in reversible DNA single-strand breaks; whereas chronic exposure causes loss of cell viability. Alcohol Alcohol 2003;38:550-8.

  • Mattson SN Schoenfeld AM Riley EP. Teratogenic effects of alcohol on brain and behavior. Alcohol Res Health 2001;25(Suppl 3):185-91.

  • Kimelberg HK Norenberg MD. Astocytes. Sci Am 1989;260:66-76.

  • Di Monte DA Royland JE Irwin I Langston JW. Astrocytes as the site for bioactivation of neurotoxins. Neurotoxicology 1996;17:697-703.

  • Ledig M Megias-Megias L Tholey G. Maternal alcohol exposure before and during pregnancy: effect on development of neurons and glial cells in culture. Alcohol Alcohol 1991;26:169-76.

  • Ledig M Tholey G Megias-Megias L Kopp P Wedler F. Combined effects of ethanol and manganese on cultured neurons and glia. Neurochem Res 1991;16:591-6.

  • Holownia A Ledig M Copin JC Tholey G. The effect of ethanol on HSP70 in cultured rat glial cells and in brain areas of rat pups exposed to ethanol in utero. Neurochem Res 1995;20:875-8.

  • Holownia A Ledig M Mapoles J Ménez JF. Acetaldehyde-induced growth inhibition in cultured rat astroglial cells. Alcohol 1996;13:93-7.

  • Holownia A Ledig M Ménez JF. Ethanol-induced cell death in cultured rat astroglia. Neuro-Toxicol Teratol 1997;19:141-6.

  • Quertemont E Tambour S Bernaerts P Zimatkin SM Tirelli E. Behavioral characterization of acetaldehyde in C57BL/6J mice: locomotor hypnotic anxiolytic and amnesic effects. Psychopharmacology (Berl) 2004;177:84-92.

  • Deitrich RA. Acetaldehyde: déjà vu du jour. J Stud Alcohol 2004;65:557-72.

  • Luke Y Ethanol. In: Goldfrank LR Flomenbaum NE Lewin NA Howland MA Hoffman RS Nelson LS editors. Goldfrank's Toxicologic Emergencies. 7th ed. New York (NY): McGraw-Hill; 2002. p. 1147-61.

  • Zimatkin SM Dietrich RA. Ethanol metabolism in the brain. Addict Biol 1997;2:387-413.

  • Hunt WA. Role of acetaldehyde in the actions of ethanol on the brain - a review. Alcohol 1996;13:147-51.

  • Aragon CM Rogan F Amit Z. Studies on ethanol-brain catalase interaction: evidence for central ethanol oxidation. Alcoh Clin Exp Res 1991;15:165-9.

  • Gill K Menez JF Lucas D Deitrich RA. Enzymatic production of acetaldehyde from ethanol in rat brain tissue. Alcohol Clin Exp Res 1992;16:910-5.

  • Aragon CM Amit Z. Differences in ethanol-induced behaviors in normal and acatalasemic mice: systematic examination using a biobehavioral approach. Pharmacol Biochem Behav 1993;44:547-54.

  • Zimatkin SM Buben AL. Ethanol oxidation in the living brain. Alcohol Alcohol 2007;42:529-32.

  • Zimatkin SM Lindros KO. Distribution of catalase in rat brain: aminergic neurons as possible targets for ethanol effects. Alcohol Alcohol 1996;31:167-74.

  • Quertemont E Eriksson CJ Zimatkin SM Pronko PS Diana M Pisano M Rodd ZA Bell RR Ward RJ. Is ethanol a prodrug? Acetaldehyde contribution to brain ethanol effects. Alcohol Clin Exp Res 2005;29:1514-21.

  • Aragon CM Spivak K Amit Z. Effect of 3-amino-124-triazole on ethanol-induced narcosis lethality and hypothermia in rats. Pharmacol Biochem Behav 1991;39:55-9.

  • Aragon CM Amit Z. The effect of 3-amino-124-triazole on voluntary ethanol consumption: evidence for brain catalase involvement in the mechanism of action. Neuropharmacology 1992;31:709-12.

  • Aragon CM Spivak K Amit Z. Blockade of ethanol induced conditioned taste aversion by 3-amino-124-triazole: evidence for catalase mediated synthesis of acetaldehyde in rat brain. Life Sci 1985;37:2077-84.

  • Tampier L Quintanilla ME. Effect of 3-amino-124-triazole on the hypothermic effect of ethanol and on ethanol tolerance development. Alcohol 1991;8:279-82.

  • Wilson JX. Antioxidant defense of the brain: a role for astrocytes. Can J Pharmacol 1997;75:1149-63.

  • Pentreath VW Slamon ND. Astrocyte phenotype and prevention against oxidative damage in neurotoxicity. Hum Exp Toxicol 2000;19:641-9.

  • Lipnik-Stangelj M Carman-Krzan M. Activation of histamine H1-receptor enhances neurotrophic factor secretion from cultured astrocytes. Inflamm Res 2004;53:245-52.

  • Uliasz TF Hewett SJ. A microtiter trypan blue absorbance assay for the quantitative determination of excitotoxic neuronal injury in cell culture. J Neurosci Methods 2000;100:157-63.

  • Protocol online. Your lab's reference book [displayed 21 April 2009]. Available at http://www.protocol-online.org/prot/Image_Techniques/Microscopy/Light_Microscopy/index.html

  • Bradford M. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-54.

  • Forn-Frías C Sanchis-Segura C. The possible role of acetaldehyde in the brain damage caused by the chronic consumption of alcohol. Rev Neurol 2003;37:485-93.

  • Davies DL Cox WE. Delayed growth and maturation of astrocytic cultures following exposure to ethanol: electron microscopic observations. Brain Res 1991;546:53-61.

  • Guizzeti M Costa LG. Inhibition of muscarinic receptor-stimulated glial cell proliferation by ethanol. J Neurochem 1996;67:2236-45.

  • Russo A Palumbo M Scifo C Cardile V Barcellona mL Renis M. Ethanol-induced oxidative stress in rat astrocytes: role of hsp70. Cell Biol Toxicol 2001;17:153-68.

  • Muscoli C Fresta M Cardile V Palumbo M Renis M Puglisi G Paolino D Nisticò S Rotiroti D Mollace V. Ethanol-induced injury in rat primary cortical astrocytes involves oxidative stress: effect of idebenone. Neurosci Lett 2002;329:21-4.

  • Gonthier B Signorini-Allibe N Soubeyran A Eysseric H Lamarche F Barret L. Ethanol can modify the effects of certain free radical-generating systems on astrocytes. Alcohol Clin Exp Res 2004;28:526-34.

  • Signorini-Allibe N Gonthier B Lamarche F Eysseric H Barret L. Chronic consumption of ethanol leads to substantial cell damage in cultured rat astrocytes in conditions promoting acetaldehyde accumulation. Alcohol Alcohol 2004;40:163-71.

  • Rottenberg H. Membrane solubility of ethanol in chronic alcoholism. The effect of ethanol feeding and its withdrawal on the protection by alcohol of rat red blood cells from hypotonic hemolysis. Biochim Biophys Acta 1986;855:211-22.

  • O'Callaghan JP Jensen KF Miller DB. Quantitative aspects of drug and toxicant induced astrogliosis. Neurochem Int 1995;26:115-24.

  • Wilson JX. Antioxidant defense of the brain: a role for astrocytes. Can J Physiol Pharmacol 1997;75:1149-63.

  • Watts LT Rathinam ML Schenker S Henderson GI. Astrocytes protect neurons from ethanol-induced oxidative stress and apoptotic death. J Neurosci Res 2005;80:655-66.

  • Poisindex [CD-ROM]. Greenwood Village USA: Thompson Micromedex; 2008.

  • Labourdette G Sensenbrenner M. Growth factors and their receptors in the central nervous system. In: Kettenmann H Ranson BR editors. Neuroglia. New York (NY): Oxford University Press; 1995. p. 441-59.

  • Lauder JM. Neurotransmitters as growth regulatory signals: role of receptors and second messengers. Trends Neurosci 1993;16:233-40.

  • Resnicoff M Rubini M Baserga R Rubin R. Ethanol inhibits insulin-like growth factor-1-mediated signalling and proliferation of C6 rat glioblastoma cells. Lab Invest 1994;71:657-62.

  • Luo J Miller MW. Growth factor-mediated neural proliferation: target of ethanol toxicity. Brain Res Brain Res Rev 1998;27:157-67.

  • Guerri C Sáez R Sancho-Tello M Martin de Aquilera E Renau-Piqueras J. Ethanol alters astrocyte development: a study of critical periods using primary cultures. Neurochem Res 1990;15:559-65.

  • Holownia A Ledig M Braszko JJ Ménez JF. Acetaldehyde cytotoxicity in cultured rat astrocytes. Brain Res 1999;833:202-8.

  • Eysseric H Gonthier B Soubeyran A Richard MJ Daveloose D Barret L. Effects of chronic ethanol exposure on acetaldehyde and free radical production by astrocytes in culture. Alcohol 2000;21:117-25.

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