Tumor necrosis factor alpha (TNF-alpha)-induced RANTES chemokine expression via activation of NF-kappaB and p38 MAP kinase: roles of TNF-alpha in alcoholic liver diseases.
OBJECTIVE
Increased concentration of plasma tumor necrosis factor alpha (TNF-alpha) correlates with the clinical course of alcoholic liver diseases. In addition, hepatic RANTES which migrates CD4 T lymphocytes to liver is increased in patients with alcoholic hepatitis. We investigated that roles of TNF-alpha on RANTES expression in hepatocytes.


METHODS
HLE cells were treated with TNF-alpha in the presence, or absence of several inhibitors. Enzyme-linked immunoassay and reverse transcriptase-polymerase chain reaction were performed for the measurement of protein production and mRNA of RANTES, respectively. Moreover, DNA-binding activity of NF-kappaB was investigated using electrophoretic mobility shift assay. To examine effects of TNF-alpha on RANTES gene expression, luciferase assay was performed.


RESULTS
TNF-alpha clearly up-regulated RANTES expression in a time-dependent fashion and induced DNA-binding activity of NF-kappaB. Moreover, TNF-alpha-induced RANTES expression was completely inhibited by SB203580, but not calphostin C and wortmannin. Luciferase assay showed that TNF-alpha increased RANTES gene expression and mutation of NF-kappaB binding sites in the RANTES promoter ablated TNF-alpha inducibility.


CONCLUSIONS
We showed that RANTES was transcriptionally induced in human hepatoma cells by treatment with TNF-alpha via activation of NF-kappaB and p38 MAP kinase, presumably suggesting that TNF-alpha-induced expression of RANTES plays important roles in cell-mediated liver injury in alcoholic liver diseases.
Hirano
Fuminori
F
Second Department of Internal Medicine, Asahikawa Medical College, Midorigaoka-higashi 2-1, Asahikawa, 078-8510, Japan. fhirano@asahikawa-med.ac.jp
Komura
Keiji
K
Fukawa
Etsushi
E
Makino
Isao
I
D000941:Antigens / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D018946:Chemokine CCL5 / Q000235:genetics*; D015870:Gene Expression / Q000187:drug effects / Q000276:immunology; D006801:Humans; D008108:Liver Diseases, Alcoholic / Q000276:immunology* / Q000378:metabolism / Q000503:physiopathology; D008113:Liver Neoplasms; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000502:physiology; D015533:Transcriptional Activation / Q000187:drug effects / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D048051:p38 Mitogen-Activated Protein Kinases
D000941:Antigens; D000970:Antineoplastic Agents; D018946:Chemokine CCL5; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases