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Background/Aims: Increased concentration of plasma TNF-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 -on RANTES expression in hepatocytes.Methods: HLE cells were treated with TNF-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-B was investigated using electrophoretic mobility shift assay. To exam effects of TNF-on RANTES gene expression, luciferase assay was performed. Results: TNF-clearly up-regulated RANTES expression in a time-dependent fashion andinduced DNA-binding activity of NF-B. Moreover, TNF--induced RANTES expression was completely inhibited by SB203580, but not calphostin C and wortmannin. Luciferase assay showed that TNF-increased RANTES gene expression and mutation of NF-B binding sites in the RANTES promoter ablated TNF-inducibility. Conclusions:We presented that RANTES was transcriptionally induced in human hepatoma cells by treatment with TNF-via activation of NF-B and p38 MAP kinase, presumably suggesting that TNF--induced expression of RANTES plays important roles in cell-mediated liver injury in alcoholic liver diseases.

Regulated upon activation, normal T-cells expressed and secreted (RANTES) mainly migrates memory type CD4+ T-lymphocytes to inflamed tissues. In this study, we examined effects of bile acids on RANTES gene expression in human hepatoma cells. Upon stimulation with hydrophobic bile acids, RANTES proteins were clearly increased. Semiquantitative RT-PCR analysis revealed that chenodeoxycholic acid (CDCA) induced RANTES mRNA expression. Moreover, RANTES was transcriptionally induced in two hepatoma cell lines by CDCA, presumably via its cognate NF-kappaB binding sites in the RANTES promoter. Electrophoretic mobility shift assay revealed that hydrophobic bile acids induced DNA-binding activity of NF-kappaB. Additionally, the magnitude of inducibility was closely associated with the hydrophobicity of bile acids. In conclusion, we might indicate that bile acids induced RANTES gene expression in human hepatoma cells, possibly suggesting that bile acids play an important role in migration of inflammatory cells by RANTES to the liver in patients with primary biliary cirrhosis.

To determine the relative potency of synthetic glucocorticoids, glucocorticoid receptor expressing cells were transfected with a hormone-inducible reporter gene, and were cultured in the presence of various glucocorticoid ligands. Hormonal inducibility was determined by means of a chloramphenicol acetyltransferase assay. Dexamethasone and prednisolone, as well as cortisol, induced the expression of the reporter gene in a dose-dependent fashion. The relative potency of each ligand was in this order when inducibility was quantitatively assessed. In conclusion, the transcription assay described here may be a convenient and alternative method to evaluate the relative potency of given glucocorticoids.