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

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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.

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Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Koshigaya Hospital, Dokkyo University School of Medicine, 2-1-50 Minami-Koshigaya, Koshigaya, Saitama, Japan

    Yasushi Shiratori PhD, Hiroshi Takada MD, Kenji Hai MD, Hiroki Kiriyama MD, Tohru Nagura MD, Mitsugu Tanaka PhD, Kazunori Matsumoto PhD & Kazuo Kamii PhD

  2. 2nd Department of Internal Medicine Faculty of Medicine, University of Tokyo, 7-3-1 Hongo, 113, Bunkyo-ku, Tokyo, Japan

    Yasushi Shiratori PhD, Hiroshi Takada MD, Kenji Hai MD, Hiroki Kiriyama MD, Tohru Nagura MD, Mitsugu Tanaka PhD, Kazunori Matsumoto PhD & Kazuo Kamii PhD

  3. Department of Pathology, Jikei University, School of Medicine, 3-25-8 Nishi-Shin'bashi, Minato-ku, Tokyo, Japan

    Yasushi Shiratori PhD, Hiroshi Takada MD, Kenji Hai MD, Hiroki Kiriyama MD, Tohru Nagura MD, Mitsugu Tanaka PhD, Kazunori Matsumoto PhD & Kazuo Kamii PhD

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  1. Yasushi Shiratori PhD
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  2. Hiroshi Takada MD
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  3. Kenji Hai MD
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  4. Hiroki Kiriyama MD
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  5. Tohru Nagura MD
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  6. Mitsugu Tanaka PhD
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  7. Kazunori Matsumoto PhD
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  8. Kazuo Kamii PhD
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Additional information

This work was supported by grants from Ministry of Education, Culture and Science in Japan: Sankyo Co; and Eisai Co.

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Shiratori, Y., Takada, H., Hai, K. et al. Generation of chemotactic factor by hepatocytes isolated from chronically ethanol-fed rats. Digest Dis Sci 37, 650–658 (1992). https://doi.org/10.1007/BF01296418

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  • DOI: https://doi.org/10.1007/BF01296418

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