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Changes in DNA strand breaks in non parenchymal cells following hepatocyte regeneration in CCl4-induced rat liver injury

  • Published:
Virchows Archiv B

Summary

DNA strand breaks (nicks) in non-parenchymal cells (NPCs) in CCl4-induced acute or chronic liver injury in rats were detected using an in situ nick translation method; their dynamic changes were analysed in relation to the proliferation pattern of hepatocytes and NPCs, as revealed by bromodeoxyuridine (BrdU)-up-take. In acute injury, hepatocyte proliferation started before centrilobular necrosis had occurred, whereas BrdU-labeled sinusoidal NPCs markedly increased only after centrilobular necrosis was apparent. DNA breakages in NPCs paralleled the proliferation pattern of these cells, suggesting that nicks are physiological, and reflect proliferation and activated gene expression. In chronic injury, liver cirrhosis developed after 9 weeks, but BrdU-labeling of hepatocytes was almost the same level as that in untreated liver. The number of BrdU-labeled NPCs showed only a slight increase, while those with DNA breakages were much more frequent in the cirrhotic stage, suggesting a significant role for NPCs in the fibrotic process. These results indicate that DNA strand breaks in NPCs act as a marker for activation states such as proliferation, differentiation and/or activated gene expression.

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References

  • Braun L, Mead JE, Panzica M, Mikumo R, Bell GI, Fausto N (1988) Transforming growth factor beta increases during liver regeneration: a possible paracrine mechanism of growth regulation. Proc Natl Acad Sci USA 85:1539–1543

    Article  PubMed  CAS  Google Scholar 

  • Collins JM (1977) Deoxyribonucleic acid structure in human diploid fibroblasts stimulated to proliferate. J Biol Chem 252:141–147

    PubMed  CAS  Google Scholar 

  • Czaja MJ, Weiner FR, Flanders KC, Giambrone M-A, Wind R, Biempica L, Zern MA (1989) In vitro and in vivo association of transforming growth factor-β1 with hepatic fibrosis. J Cell Biol 108:2477–2482

    Article  PubMed  CAS  Google Scholar 

  • Dawson BA, Lough J (1988) Immunocytochemical localization of transient DNA strand breaks in differentiating myotubes using in situ nick translation. Dev Biol 127:362–367

    Article  PubMed  CAS  Google Scholar 

  • Fausto N, Mead JE (1989) Regulation of liver growth: protooncogenes and transforming growth factors. Lab Invest 60:4–13

    PubMed  CAS  Google Scholar 

  • Friedman SL (1990) Cellular sources of collagen and regulation of collagen production in liver. Sem Liv Dis 10:20–29

    CAS  Google Scholar 

  • Geerts A, Schellinck P, Bouwens L, Wisse E (1988) Cell population kinetics of Kuppfer cells during the onset of fibrosis in rat liver by chronic carbon tetrachloride administration. J Hepatol 6:50–56

    Article  PubMed  CAS  Google Scholar 

  • Gratzner HG (1982) Monoclonal antibody to 5-bromo-and 5-iododeoxy-uridine: a new reagent for detection of DNA replication. Science 218:474–476

    Article  PubMed  CAS  Google Scholar 

  • Herbst H, Milani S, Schuppan D, Stein H (1991) Temporal and spatial patterns of proto-oncogene expression at early stages of toxic liver injury in the rat. Lab Invest 65:324–333

    PubMed  CAS  Google Scholar 

  • Iseki S (1986) DNA strand breaks in rat tissues as detected by in situ nick translation. Exp Cell Res 167:311–326

    Article  PubMed  CAS  Google Scholar 

  • Katsumoto F, Miyazaki K, Nakayama F (1989) Stimulation of DNA synthesis in hepatocytes by Kupffer cells after partial hepatectomy. Hepatology 9:405–410

    Article  PubMed  CAS  Google Scholar 

  • Kent G, Volini FI, Minick OT, Orfei E, Dela Huerga J (1964) Effect of iron loading upon the formation of collagen in the hepatic injury induced by carbon tetrachloride. Am J Pathol 45:129–155

    PubMed  CAS  Google Scholar 

  • Kinoshita T, Tashiro T, Nakamura T (1989) Marked increase of HGF mRNA in non-parenchymal liver cells of rats treated with hepatotoxins. Biochem Biophys Res Comm 165:1229–1234

    Article  PubMed  CAS  Google Scholar 

  • Lough J, Rosenthall L, Arzoumanian A, Goresky CA (1987) Kupffer cell depletion associated with capillarization of liver sinusoids in carbon tetrachloride-induced rat liver cirrhosis. J Hepatol 5:190–198

    Article  PubMed  CAS  Google Scholar 

  • Maehara Y, Anai H, Kusumoto T, Sakaguchi Y, Sugimachi K (1989) Nick translation detection in situ of cellular DNA strand break induced by irradiation. Am J Pathol 134:7–10

    PubMed  CAS  Google Scholar 

  • Maher JJ, McGuire RF (1990) Extracellular matrix gene expression increases preferentially in rat lipocytes and sinusoidal endothelial cells during hepatic fibrosis in vivo. J Clin Invest 86:1641–1648

    Article  PubMed  CAS  Google Scholar 

  • Milani S, Herbst H, Schuppan D, Hahn EG, Stein H (1989) In situ hybridization for procollagen types I, III and IV mRNA in normal and fibrotic rat liver: evidence for predominant expression in non-parenchymal liver cells. Hepatology 10:84–92

    Article  PubMed  CAS  Google Scholar 

  • Moran R, Darzynkiewicz Z, Staiano-coico L, Melamed MR (1985) Detection of 5-bromodeoxyuridine (BrdUrd) incorporation by monoclonal antibodies: role of the DNA denaturation step. J Histochem Cytochem 33:821–827

    PubMed  CAS  Google Scholar 

  • Nakamura T, Yoshimoto K, Nakayama F, Tomita Y, Ichihara A (1983) Reciprocal modulation of growth and differentiated functions of mature rat hepatocytes in primary culture by cell-cell contact and cell membranes. Proc Natl Acad Sci USA 80:7229–7233

    Article  PubMed  CAS  Google Scholar 

  • Nakamura T, Arakaki R, Ichihara A (1988) Interleukin-1β is a potent growth inhibitor of adult rat hepatocytes in primary culture. Exp Cell Res 179:488–497

    Article  PubMed  CAS  Google Scholar 

  • Nakatsukasa H, Evarts RP, Hsia C-C, Thorgeirsson SS (1990) Transforming growth factor-beta 1 and type I procollagen transcripts during regeneration and early fibrosis of rat liver. Lab Invest 63:171–180

    PubMed  CAS  Google Scholar 

  • Nostrant TT, Miller DL, Appelman HD, Gumucio JJ (1978) Acinar distribution of liver cell regeneration after selective zonal injury in the rat. Gastroenterology 75:181–186

    PubMed  CAS  Google Scholar 

  • Ogawa K, Suzuki J, Mukai H, Mori M (1985) Healing of focal injury in rat liver. Am J Pathol 119:158–167

    PubMed  CAS  Google Scholar 

  • Panduro A, Shalaby F, Biempica L, Shafritz DA (1988) Changes in albumin, alpha-fetoprotein and collagen gene transcription in CCl4-induced hepatic fibrosis. Hepatology 8:259–266

    Article  PubMed  CAS  Google Scholar 

  • Ramadori G (1991) The stellate cell (Ito cell, fat-storing cell, lipocyte, perisinusoidal cell) of the liver. Virchows Arch [B] 61:147–158

    Article  CAS  Google Scholar 

  • Saez JC, Bennett MVL, Spray DC (1987) Carbon tetrachloride at hepatotoxic levels blocks reversibly gap junctions between rat hepatocytes. Science 236:967–969

    Article  PubMed  CAS  Google Scholar 

  • Sasaki Y, Hayashi N, Morita Y, Ito T, Kasahara A, Fusamoto H, Sato N, Tohyama M, Kamada T (1989) Cellular analysis of c-Ha-ras gene expression in rat liver after CCl4 administration. Hepatology 10:494–500

    Article  PubMed  CAS  Google Scholar 

  • Scarpelli DG, Iannaccone PM (1990) Cell injury and errors of metabolism. In: Kissane JM (ed) Anderson’s Pathology (9th ed). C.V. Mosby, St. Louis, pp 16–18

    Google Scholar 

  • Strain AJ, Frazer A, Hill DJ, Milner RDG (1987) Transforming growth factor beta inhibits DNA synthesis in hepatocytes isolated from normal and regenerating rat liver. Biochem Biophys Res Comm 145:436–442

    Article  PubMed  CAS  Google Scholar 

  • Tournier I, Legres L, Shoevaert D, Feldmann G, Bernuau D (1988) Cellular analysis of alpha-fetoprotein gene activation during carbon tetrachloride and D-galactosamine-induced acute liver injury in rats. Lab Invest 59:657–665

    PubMed  CAS  Google Scholar 

  • Ueda K, Nakata M, Urata Y, Konishi E, Itoi H, Ashihara T (1990) Detection of DNA injury by in situ nick translation. Effect of fixation and air-drying on nuclear DNA. Igakunoayumi 152:247–248

    Google Scholar 

  • Unanue ER (1976) Secretory function of mononuclear phagocytes. Am J Pathol 83:396–417

    PubMed  CAS  Google Scholar 

  • van Eyken P, Sciot R, Desmet VJ (1990) Expression of novel extracellular matrix component in normal and diseased human liver. An immunohistochemical study. J Hepatol 11:45–52

    Google Scholar 

  • Yokoi Y, Namihisa T, Matsuzaki K Miyazaki A, Yamaguchi Y (1988) Distribution of Ito cells in experimental hepatic fibrosis. Liver 8:48–52

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Pathology, Shinshu University School of Medicine, 3-1-1 Asahi, 390, Matsumoto, Japan

    Toshitsugu Nakamura & Masao Hotchi

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  1. Toshitsugu Nakamura
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  2. Masao Hotchi
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Nakamura, T., Hotchi, M. Changes in DNA strand breaks in non parenchymal cells following hepatocyte regeneration in CCl4-induced rat liver injury. Virchows Archiv B Cell Pathol 63, 11–16 (1993). https://doi.org/10.1007/BF02899239

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  • DOI: https://doi.org/10.1007/BF02899239

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