1992 Fiscal Year Final Research Report Summary
A MECHANISM OF INFILTRATION OF POLYMORPHONUCLEAR CELLS AND KUPFFER CELLS IN ALCOHOLIC LIVER INJURY : PRODUCTION OF CHEMOTACTIC FACTOR BY HEPATOCYTES EXPOSED TO ETHANOL
| Project/Area Number |
03670376
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Gastroenterology
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| Research Institution | DOKKYO UNIVERSITY SCHOOL OF MEDICINE |
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Principal Investigator |
MATSUMOTO Kazunori DOKKYO UNIVERSITY, SCHOOL OF MEDICINE, ASSOCIATE PROFESSOR, 医学部, 助教授 (20010499)
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| Co-Investigator(Kenkyū-buntansha) |
SHIRATORI Yasushi UNIVERSITY OF TOKYO, FACULTY OF MEDICINE, 医学部, 助手 (70196624)
TAKADA Hiroshi DOKKYO UNIVERSITY, SCHOOL OF MEDICINE, 医学部, 助手 (50201596)
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| Project Period (FY) |
1991 – 1992
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| Keywords | Chemotactic factor / Isolated hepatocytes / Polymorphonuclear cells / Kupffer cells / Ethanol / Superoxide anion / Ethanol-fed rat / Alcoholic liver injury |
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| Research Abstract |
Ethanol induces acute hepatic inflammation histologically characterized by focal necrosis of hepatocytes infiltrated with polymorphonuclear cells (PMNs). To know the mechanism and pathogenesis of cellular infiltration in alcoholic liver injury, we investigated a novel chemotactic factor produced by hepatocytes exposed to ethanol. Hepatocytes and Kupffer cells were isolated from rat liver by perfusion and digestion with collagenase followed by differential centrifugation on metrizamide gradient. Rat PMNs were prepared from blood by dextran sedimentation and Hypaque-Ficoll gradient centrifugation.
When hepatocytes isolated from normal rat liver were cultured with ethanol (30-90mM), chemotactic activity for rat PMNs and Kupffer cells was demonstrated in the culture supernatant. Similar results were obtained when hepatocytes isolated from chronically ethanol-fed rats were cultured in vitro. Freeze-thawing or dialysis of the culture medium gave little effect on chemotactic activity, while trypsin digestion and heat (56ﾟC, 30min) markedly reduced the activity. Addition of inhibitor of transcription or protein synthesis to the culture medium decreased production of the chemotactic factor from hepatocytes. Chemotactic activity was revealed in two fractions at molecular weight of 20-25 k daltons and 40-50 k daltons by use of gel chromatography with Sephadex G-150. Production of superoxide anion by PMNs and Kupffer cells was enhanced in the presence of the chemotactic factor.
These results suggested that accumulation and activation of PMNs and Kupffer cells in the liver were induced by the chemotactic factor and that this chemotactic factor might contribute to the pathogenesis of alcoholic liver injury.
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