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Digestive Diseases and Sciences

, Volume 37, Issue 5, pp 650–658 | Cite as

Generation of chemotactic factor by hepatocytes isolated from chronically ethanol-fed rats

  • Yasushi Shiratori
  • Hiroshi Takada
  • Kenji Hai
  • Hiroki Kiriyama
  • Tohru Nagura
  • Mitsugu Tanaka
  • Kazunori Matsumoto
  • Kazuo Kamii
Original Articles

Abstract

In an attempt to clarify a mechanism of polymorphonuclear cell and/or macrophage infiltration in alcoholic liver disease, we investigated a novel chemotactic and activating factor generated by rat hepatocytes isolated from the chronically ethanol-fed rats. Hepatocytes and hepatic macrophages were isolated from rat liver by perfusion and digestion with collagenase and subsequently by differential centrifugation on a metrizamide gradient. Rat polymorphonuclear cells were prepared from blood by the dextran sedimentation and Hypaque-Ficoll technique. Chemotactic activity was measured as migration of polymorphonuclear cells or hepatic macrophages using a chemotactic chamber. When hepatocytes isolated from the ethanol-fed rats were culturedin vitro, chemotactic activity for rat polymorphonuclear cells and hepatic macrophages was demonstrated in the culture supernatant. Inhibitors of transcription and protein synthesis reduced generation of chemotactic factor from these hepatocytes. Chemotactic activity of the conditioned medium was reduced after trypsin (0.25%, 37° C, 30 min) or heat (56° C, 30 min) treatment. The chemotactic activity was eluted at molecular weights of 20–25 kDa and 40–45 kDa following Sephadex G-150 chromatography. Superoxide anion production by polymorphonuclear cells and hepatic macrophages under the stimulation of phorbolmyristate acetate was enhanced in the presence of this chemotactic factor. This chemotactic factor may contribute to the pathogenesis of alcoholic liver disease.

Key Words

ethanol diets hepatocytes chemotactic factor polymorphonuclear cells Kupffer cells chemotaxis 

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References

  1. 1.
    International Group (MacSween RNM, et al): Alcoholic liver disease: Morphological manifestations. Lancet i:707–711, 1981Google Scholar
  2. 2.
    Takahashi T, Kamimura T, Ichida F: Ultrastructural findings on polymorphonuclear leucocyte infiltration and acute hepatocellular damage in alcoholic hepatitis. Liver 7:347–358, 1987Google Scholar
  3. 3.
    Peters M, Liebman HA, Tong MJ, Tinberg HM: Alcoholic hepatitis: Granulocyte chemotactic factor from Mallory body-stimulated human peripheral blood mononuclear cells. Clin Immunol Immunopathol 28:418–430, 1983Google Scholar
  4. 4.
    Perez HD, Roll FJ, Bissell DM, Shak S, Goldstein IM: Production of chemotactic activity for polymorphonuclear leukocytes by cultured rat hepatocytes exposed to ethanol. J Clin Invest 74:1350–1357, 1984Google Scholar
  5. 5.
    Roll FJ, Bissell DM, Perez HD: Human hepatocytes metabolizing ethanol generate a non-polar chemotactic factor for human neutrophils. Biochem Biophys Res Commun 137:688–694, 1986Google Scholar
  6. 6.
    Roll FJ, Alexander M, Perez HD: Generation of chemotactic activity for neutrophils by liver cells metabolizing ethanol. Free Rad Biol Med 7:549–555, 1989Google Scholar
  7. 7.
    Lautenschlager I, Vaananen H, Kulonen E: Qualitative study on the Kupffer cells in the liver of ethanol- and carbon tetrachloride-treated rats. Acta Pathol Microbiol Immunol Scand Sect C 90:347–351, 1982Google Scholar
  8. 8.
    Shiratori Y, Teraoka H, Matano S, Matsumoto K, Kamii K, Tanaka M: Kupffer cell function in chronic ethanol-fed rats. Liver 9:351–359, 1989Google Scholar
  9. 9.
    Popper H, Lieber CS: Histogenesis of alcoholic fibrosis and cirrhosis in the baboon. Am J Pathol 98:695–716, 1980Google Scholar
  10. 10.
    Rubin E, Lieber CS: Fatty liver, alcoholic hepatitis and cirrhosis produced by alcohol in primates. N Engl J Med 290:128–135, 1974Google Scholar
  11. 11.
    Lieber CS, DeCarli LM: Animal models of ethanol dependence and liver injury in rats and baboons. Fed Proc 35:1232–1236, 1976Google Scholar
  12. 12.
    Bhagwandeen BS, Apte M, Manwarring L, Dickeson J: Endotoxin induced hepatic necrosis in rats on an alcohol diet. J Pathol 151:47–53, 1987Google Scholar
  13. 13.
    Lieber CS, DeCarli LM, Sorrell MF: Experimental methods of ethanol administration. Hepatology 10:501–510, 1989Google Scholar
  14. 14.
    Poklis A, Mackell MA: Evaluation of a modified alcohol dehydrogenase assay for the determination of ethanol in blood. Clin Chem 28:2125–2127, 1982Google Scholar
  15. 15.
    Seglen PO: Preparation of rat liver cells. III. Enzymatic requirements for tissue dispersion. Exp Cell Res 82:391–398, 1973Google Scholar
  16. 16.
    Laskin DL, Pilaro AM: Potential role of activated macrophages in acetaminophen hepatotoxicity. I. Isolation and characterization of activated macrophages from rat liver. Toxicol Appl Pharmacol 86:204–215, 1986Google Scholar
  17. 17.
    Shiratori Y, Geerts A, Ichida T, Kawase T, Wisse E: Kupffer cells from CCl4-induced fibrotic livers stimulate proliferation of fat-storing cells. J Hepatol 3:294–303, 1986Google Scholar
  18. 18.
    Chodirker WB, Bock GN, Vaughan JH: Isolation of human PMN leukocytes and granules. Observation on early blood dilution and on heparin. J Lab Clin Med 71:9–16, 1968Google Scholar
  19. 19.
    Ferrante A, Thong YH: A rapid one-step procedure for purification of mononuclear and polymorphonuclear leukocytes from human blood using a modification of the hypaqueficoll technique. J Immunol Methods 24:389–393, 1978Google Scholar
  20. 20.
    Onozaki K, Ichikawa M, Hashimoto T: A simplified assay for neutrophil chemotactic activity of lymphocyte culture supernatants. Jpn J Exp Med 49:281–287, 1979Google Scholar
  21. 21.
    Chenoweth DE, Lane TA, Rowe JG, Hugli TE: Quantitative comparisons of neutrophil chemotaxis in four animal species. Clin Immunol Immunopathol 15:525–535, 1980Google Scholar
  22. 22.
    Weiss RE, Reddi AH: Isolation and characterization of rat plasma fibronectin. Biochem J 197:529–534, 1981Google Scholar
  23. 23.
    Shiratori Y, Kawase T, Shiina S, Okano K, Sugimoto T, Teraoka H, Matano S, Matsumoto K, Kamii K: Modulation of hepatotoxicity by macrophages in the liver. Hepatology 8:815–821, 1988Google Scholar
  24. 24.
    Lue SL, Paronetto F, Lieber CS: Cytoxicity of mononuclear cells and vulnerability of hepatocytes in alcoholic fatty liver of baboons. Liver 1:264–267, 1981Google Scholar
  25. 25.
    Mavier P, Preaux AM, Guigui B, Lescs MC, Zafrani ES, Dhumeaux D:In vitro toxicity of polymorphonuclear neutrophils to rat hepatocytes: Evidence for a proteinase-mediated mechanism. Hepatology 8:254–258, 1988Google Scholar
  26. 26.
    Synderman R, Mergenhagen SE: Chemotaxis of macrophages.In Immunobiology of the Macrophages. DS Nelson (ed). New York, Academic Press, 1976, pp 323–348Google Scholar
  27. 27.
    Wilkinson PC: Cellular and molecular aspects of chemotaxis of macrophages and monocytes.In Immunobiology of the Macrophages. DS Nelson (ed). New York, Academic Press, 1976, pp 349–365Google Scholar
  28. 28.
    Boyden S: The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes. J Exp Med 115:453–466, 1962Google Scholar
  29. 29.
    Keller HU, Sorkin E: Chemotaxis of leucocytes. Experientia 24:641–652, 1968Google Scholar
  30. 30.
    Tran-Thi TA, Phillips J, Falk H, Decker K: Toxicity ofd-galactosamine for rat hepatocytes in monolayer culture. Exp Mol Pathol 42:89–116, 1985Google Scholar
  31. 31.
    Tanner A, Keyhani A, Reiner R, Holdstock G, Wright R: Proteolytic enzymes released by liver macrophages may promote hepatic injury in a rat model of hepatic damage. Gastroenterology 80:647–654, 1981Google Scholar
  32. 32.
    Maier RV, Ulevitch RJ: The response of isolated rabbit hepatic macrophages(H-mø) to lipopolysaccharide (LPS). Circ Shock 8:165–181, 1981Google Scholar
  33. 33.
    Laskin DL, Pilaro AM, Ji S: Potential role of activated macrophages in acetaminophen hepatotoxicity. II. Mechanism of macrophage accumulation and activation. Toxicol Appl Pharmacol 86:216–226, 1986Google Scholar
  34. 34.
    Fantone JC, Ward PA: Polymorphonuclear leukocyte-mediated cell and tissue injury: Oxygen metabolites and their relations to human disease. Hum Pathol 16:973–978, 1985Google Scholar

Copyright information

© Plenum Publishing Corporation 1992

Authors and Affiliations

  • Yasushi Shiratori
    • 1
    • 2
    • 3
  • Hiroshi Takada
    • 1
    • 2
    • 3
  • Kenji Hai
    • 1
    • 2
    • 3
  • Hiroki Kiriyama
    • 1
    • 2
    • 3
  • Tohru Nagura
    • 1
    • 2
    • 3
  • Mitsugu Tanaka
    • 1
    • 2
    • 3
  • Kazunori Matsumoto
    • 1
    • 2
    • 3
  • Kazuo Kamii
    • 1
    • 2
    • 3
  1. 1.Department of Gastroenterology and Hepatology, Koshigaya HospitalDokkyo University School of MedicineKoshigaya, SaitamaJapan
  2. 2.2nd Department of Internal Medicine Faculty of MedicineUniversity of TokyoBunkyo-ku, TokyoJapan
  3. 3.Department of PathologyJikei University, School of MedicineTokyoJapan

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