ENHANCED INTERLEUKIN-8 PRODUCTION BY CULTURED CHANG LIVER-CELLS SUBJECTED TO ETHANOL EXPOSURE AND SUBSEQUENT STIMULATION WITH TUMOR-NECROSIS-FACTOR-ALPHA

Citation
H. Ohira et al., ENHANCED INTERLEUKIN-8 PRODUCTION BY CULTURED CHANG LIVER-CELLS SUBJECTED TO ETHANOL EXPOSURE AND SUBSEQUENT STIMULATION WITH TUMOR-NECROSIS-FACTOR-ALPHA, Digestion, 58(1), 1997, pp. 72-77
Citations number
26
Categorie Soggetti
Gastroenterology & Hepatology
Journal title
ISSN journal
00122823
Volume
58
Issue
1
Year of publication
1997
Pages
72 - 77
Database
ISI
SICI code
0012-2823(1997)58:1<72:EIPBCC>2.0.ZU;2-5
Abstract
We investigated the production of interleukin-8 (IL-8) and chemotactic activity released from Chang liver cells subjected to long-term treat ment with ethanol (Et) and subsequent stimulation with tumor necrosis factor-alpha (TNF-alpha). Chang liver cells were cultured in the prese nce of 5, 50 or 100 mmol/l Et for 4 weeks and then treated with recomb inant TNF-alpha (1, 10, 100 U/ml). The culture supernatants were assay ed for IL-8 using a sandwich ELISA and chemotactic activity was measur ed using a chemotactic chamber. Total RNA was also extracted from thes e cells and IL-8 mRNA was assayed by RT-PCR. In addition, TNF-receptor expression on the Et-treated cells was analyzed by flow cytometry. IL -8 levels in supernatants of cells stimulated with 100 U/ml of TNF-alp ha for 48 h rose significantly with increasing concentrations of Et an d values obtained were as follows: 4,918 +/- 244.4 pg/ml at 0 mmol/l E t, 5,335 +/- 266.2 pg/ml at 5 mmol/l Et, 8,726 +/- 873.4 pg/ml at 50 m mol/l Et and 9,134 +/- 866.0 pg/ml at 100 mmol/l Et. The chemotactic a ctivity also increased with increasing concentrations of Et and was al most completely suppressed by anti-IL-8 antibody. Using semiquantitati ve analysis of radioactivity of IL-8 mRNA using a P-32 gamma ATP-label ed primer for IL-8 mRNA, Et-treated cells were shown to have markedly higher levels of radioactivity than untreated cells. In addition, TNF- receptor expression was significantly higher in cells treated with 100 mmol/l Et. These data suggest that long-term Et treatment of Chang li ver cells stimulated with TNF-alpha may enhance transcription of the I L-8 gene with up-regulation of the TNF receptor.