Research Article| Volume 40, ISSUE 2, P219-227, February 2004

Large-scale gene profiling of the liver in a mouse model of chronic, intragastric ethanol infusion


      Background/Aims: The mechanisms underlying alcohol-induced liver injury are not fully elucidated. An approach in this direction would consist of an all-inclusive assessment of gene expression in the liver. The purpose of this study was to perform a comprehensive analysis of gene expression in the livers of mice treated with ethanol by means of intragastric infusion.
      Methods: An ethanol- or glucose-enriched liquid diet was fed to animals for 4 weeks via a long-term gastrostomy catheter. The animals were killed and plasma alanine:2-oxoglutarate aminotransferase (ALT) assay, liver histology and total RNA analysis by microarray gene technology were performed.
      Results: Alcohol increased ALT, induced steatosis, necrosis and inflammation. A total of 12,423 genes were analyzed for expression out of which 4867 were expressed by the liver. Alcohol repressed expression of 11 genes, induced expression of 13 genes, and up- or down-regulated expression of 44 and 42 genes >2-fold, respectively. Gene expression analysis identified several genes that have not previously been tested for alcohol effects.
      Conclusions: This study: (i) expands the knowledge of mechanism(s) of action of ethanol; (ii) indicates novel pathways of ethanol action on the liver, and (iii) illustrates the utility of microarray gene analysis in hepatology research.


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        • Morimoto M.
        • Hagbjörg A.-L.
        • Nanji A.A.
        • Ingelman-Sundberg M.
        • Lindros K.O.
        • Fu P.C.
        • et al.
        Role of cytochrome P450 2E1 in alcohol liver disease pathogenesis.
        Alcohol. 1993; 10: 459-464
        • Ronis M.J.J.
        • Huang J.
        • Crouch J.
        • Mercado C.
        • Irby D.
        • Valentine C.R.
        • et al.
        Cytochrome P450 CYP 2E1 induction during chronic alcohol exposure occurs by a two-step mechanism associated with blood alcohol concentrations in rats.
        J Pharmacol Exp Ther. 1993; 264: 944-950
        • Gordon A.S.
        • Collier K.
        • Diamond I.
        Ethanol regulation of adenosine receptor-stimulated cAMP levels in a cloned cell line: an invitro model of cellular tolerance to ethanol.
        Proc Natl Acad Sci USA. 1986; 83: 2105-2108
        • Hong F.
        • Kim W.H.
        • Tian Z.
        • Jaruga B.
        • Ishak E.
        • Shen X.
        • Gao B.
        Elevated interleukin-6 during ethanol consumption acts as a potential endogenous protective cytokine against ethanol-induced apoptosis in the liver: involvement of induction of Bcl-2 and Bcl-XL proteins.
        Oncogene. 2002; 21: 32-43
        • Mechly-Rosen D.
        • Chang F.-G.
        • Cheever L.
        • Kim M.
        • Diamond I.
        • Gordon A.S.
        Chronic ethanol causes heterologous desensitization of receptors by reducing ∀S messenger RNA.
        Nature (London). 1988; 333: 848-850
        • Deaciuc I.V.
        • Fortunato F.
        • D'Souza N.B.
        • Hill D.B.
        • Schmidt J.
        • Lee E.Y.
        • McClain C.J.
        Modulation of caspase-3 activity and Fas ligand mRNA expression in rat liver cells in vivo by alcohol and lipopolysaccharide.
        Alcohol Clin Exp Res. 1999; 23: 349-356
        • Deaciuc I.V.
        • D'Souza N.B.
        • de Villiers W.J.S.
        • Burikhanov R.
        • Sarphie T.G.
        • Hill D.B.
        • McClain C.J.
        Inhibition of caspases in vivo protects the rat liver against alcohol-induced sensitization to bacterial lipopolysaccharide.
        Alcohol Clin Exp Res. 2001; 25: 935-943
        • Deaciuc I.V.
        • D'Souza N.B.
        • Burikhanov R.
        • Lee E.Y.
        • Tarba C.N.
        • McClain C.J.
        • de Villiers W.J.S.
        Epidermal growth factor protects the liver against alcohol-induced injury and sensitization to bacterial lipopolysaccharide.
        Alcohol Clin Exp Res. 2002; 26: 864-874
        • Deaciuc I.V.
        • Fortunato F.
        • D'Souza N.B.
        • Hill D.B.
        • McClain C.J.
        Chronic alcohol exposure of rats exacerbates apoptosis in hepatocytes and sinusoidal endothelial cells.
        Hepatol Res. 2002; 19: 306-324
        • Koteish A.
        • Yang S.Q.
        • Lin H.Z.
        • Huand X.
        • Diehl A.M.
        Chronic ethanol exposure potentiates lipopolysaccharide liver injury despite inhibiting N-terminal kinase and caspase-3 activation.
        J Biol Chem. 2002; 277: 13037-13044
        • Yacoub L.K.
        • Fogt F.
        • Griniuviene B.
        • Nanji A.A.
        Apoptosis and Bcl-2 protein expression in experimental alcoholic liver disease in the rat.
        Alcohol Clin Exp Res. 1995; 19: 854-859
        • Nieto N.
        • Greenwel P.
        • Friedman S.L.
        • Zhang F.
        • Dannenberg A.J.
        • Cederbaum A.I.
        Ethanol and arachidonic acid increase 2(I) collagen expression in rat hepatic stellate cells overexpressing cytochrome P450 2E1. Role of H2O2 and cyclooxygenase-2.
        J Biol Chem. 2000; 275: 20136-20145
        • Takase S.
        • Takada A.
        • Urashima S.
        • Enyama K.
        • Fujii M.
        Changes in the markers related to collagen synthesis in the liver of chronically alcohol-treated rats.
        Gastroenterol Jpn. 1992; 27: 54-60
        • Zern M.A.
        • Leo M.A.
        • Giambrone M.A.
        • Lieber C.S.
        Increased type I procollagen mRNA levels and in vitro protein synthesis in the baboon model of chronic alcoholic liver disease.
        Gastroenterology. 1985; 89: 1123-1131
        • Deaciuc I.V.
        Alcohol and cytokine networks.
        Alcohol. 1997; 14: 421-430
        • Lu S.C.
        • Huang Z.Z.
        • Yang J.M.
        • Tsukamoto H.
        Effect of ethanol and high-fat feeding on hepatic γ-glutamylcysteine synthetase subunit expression in the rat.
        Hepatology. 1999; 30: 209-214
        • Cassini A.
        • Galli G.
        • Salazano R.
        • Ceni E.
        • Francheschelli F.
        • Rotella C.M.
        • Surrenti C.
        Acetaldehyde induces c-fos and c-jun proto-oncogenes in fat storing cells culture through protein kinase C activation.
        Alcohol Alcohol. 1994; 29: 303-314
        • Bulera S.J.
        • Eddy S.M.
        • Ferguson E.
        • Jatkoe T.A.
        • Reindel J.F.
        • Bleavins M.R.
        • De La Iglesia F.A.
        RNA expression in the early characterization of hepatotoxicants in Wistar rats by high-density DNA microarrays.
        Hepatology. 2002; 33: 1239-1258
        • Hansen T.
        • Dangers M.
        • Borlak J.
        • Gerstmayer B.
        • Tomiuk S.
        • Hofmann K.
        • Hübel F.
        • et al.
        Compound profiling using cDNA array based differential expression of human and rat hepatocytes.
        Toxicol Sci. 2002; 66 (abstract 45): 9-10
        • Harries H.M.
        • Fletcher S.T.
        • Duggan C.M.
        • Baker V.A.
        The use of genomics technology to investigate gene expression changes in cultured human liver cells.
        Toxicol In Vitro. 2001; 15: 309-405
        • Hite R.
        • Bowman M.
        • Asileti U.
        • Patel V.
        • Smith R.
        • Polavarapu R.
        Chronic alcohol induced tissue injury: microarray analyses of differential gene regulation.
        Fed Am Soc Exp Biol J. 2002; 16 (Abstract 16713)
        • Shirota Y.
        • Kaneko S.
        • Honda M.
        • Kawai H.F.
        • Kobayashi K.
        Identification of differentially expressed genes in hepatocellular carcinoma with cDNA microarrays.
        Hepatology. 2001; 33: 832-840
        • Shakel N.A.
        • Gorrell M.D.
        • McCaughan G.W.
        Gene array analysis and the liver.
        Hepatology. 2002; 36: 1313-1325
        • Tsukamoto H.
        • Reidelberger R.D.
        • French S.W.
        • Largman C.
        Long-term cannulation model for blood sampling and intragastric infusion in the rat.
        Am J Physiol. 1984; 247: R595-R599
        • Järvelainen H.A.
        • Ingelman-Sundberg M.
        • Lindros K.O.
        Effect of chronic coadministration of endotoxin and ethanol on rat liver pathology and proinflammatory and antiinflammatory cytokines.
        Hepatology. 1999; 29: 1503-1510
        • Chomczinski P.
        • Sacchi N.
        Single step method of RNA isolation by acid guanidine thiocyanate–phenol–chloroform extraction.
        Anal Biochem. 1987; 62: 156-161
        • Benjamini Y.
        • Hochberg Y.
        Controlling the false discovery rate: a practical and powerful approach to multiple testing.
        J R Stat Soc Ser B. 1995; 57: 289-300
        • French S.W.
        Intragastric ethanol infusion model for cellular and molecular studies of alcoholic liver disease.
        J Biomed Sci. 2001; 8: 20-27
        • Uesugi T.
        • Froh M.
        • Arteel G.E.
        • Bradford B.U.
        • Wheeler M.D.
        • Gäbele E.
        • Isayama F.
        • Thurman R.G.
        Role of lipopolysaccharide-binding protein in early alchol-induced liver injury in mice.
        J Immunol. 2002; 168: 2963-2969
        • Yin M.
        • Wheeler M.D.
        • Kono H.
        • Bradford B.U.
        • Galucci R.M.
        • Luster M.I.
        • Thurman R.G.
        Essential role of tumor necrosis factor α in EtOH-induced liver injury in mice.
        Gastroenterology. 1999; 117: 942-952
        • Watkins P.B.
        Role of cytochromes P450 in drug metabolism and hepatotoxicity.
        Semin Liv Dis. 1990; 10: 235-250
        • Dalke D.D.
        • Sorrell M.F.
        • Casey C.A.
        • Tuma D.J.
        Chronic ethanol administration impairs receptor- mediated endocytosis of epidermal growth factor by rat hepatocytes.
        Hepatology. 1990; 12: 1085-1091
        • Brindley D.N.
        What factors control hepatic triacylglycerol accumulation in alcohol abuse?.
        Biochem Soc Trans. 1998; 16: 251-253
        • Polokoff M.A.
        • Simon T.J.
        • Harris R.A.
        • Simon F.R.
        • Iwahashi M.
        Chronic ethanol increases liver plasma membrane fluidity.
        Biochemistry. 1985; 24: 3114-3120
        • Fernandez-Checa J.C.
        • Ookhtens M.
        • Kaplowitz N.
        Effects of chronic ethanol feeding on rat hepatocytic glutathione compartmentation, efflux and response to incubation with ethanol.
        J Clin Invest. 1987; 80: 57-62
        • Hirano T.
        • Kaplowitz N.
        • Tsukamoto H.
        • Kamimura S.
        • Fernandez-Checa J.C.
        Hepatic mitochondrial glutathione depletion and progression of experimental alcoholic liver disease in rats.
        Hepatology. 1992; 16: 1423-1427
        • Tomita M.
        • Itoh H.
        • Ishikawa N.
        • Higa A.
        • Ide H.
        • Murakumo Y.
        • et al.
        Molecular cloning of mouse intestinal trefoil factor and its expression during goblet cell change.
        Biochem J. 1995; 311: 293-297
        • Rodriguez S.
        • Van Aken E.
        • Van Bocx S.
        • Attoub S.
        • Nguyen Q.D.
        • Westley B.R.
        • et al.
        Trefoil peptides as proangiogenic factors in vivo and in vitro: implication of cyclooxygenase-2 and EGF receptor signaling.
        Fed Am Soc Exp Biol J. 2003; 17: 7-16
        • Hoffmann F.
        • Jagla W.
        • Wiede A.
        Molecular medicine of TFF-peptides: from gut to brain.
        Histol Histopathol. 2001; 16: 319-334
        • Nolan J.P.
        • Camara D.S.
        Intestinal endotoxins and macrophages as mediators of liver injury.
        Trans Am Clin Chem Assoc. 1988; 100: 115-125
        • Thurman R.G.
        Mechanisms of hepatic toxicity. II. Alcoholic liver injury involves activation of Kupffer cells by endotoxin.
        Am J Physiol. 1998; 275: G605-G611
        • Chang L.M.S.
        • Bollum F.J.
        Deoxynucleotide-polymerizing enzymes of calf thymus gland. V. Homogeneous terminal deoxynucleotidyl transferase.
        J Biol Chem. 1971; 246: 909-916
        • Aroca P.
        • Garcia-Borron J.C.
        • Solano F.
        • Lozano J.A.
        Regulation of mammalian melanogenesis I: partial purification and characterization of a dopachrom converting factor: dopachrome tautomerase.
        Biochim Biophys Acta. 1990; 1035: 266-275