Regulation of NF-kappaB and ICAM-1 expression in human airway epithelial cells.
The aim of this study was to elucidate the redox regulation of cytokine-induced NF-kappaB activation and NF-kappaB mediated gene induction in A549 cells and primary cultures of human airway epithelial cells. In A549 cells, Western blot analysis showed transient depletion of IkappaBalpha after 15 min IL-1beta treatment followed by its reappearance after 60 min, indicating efficient NF-kappaB-driven gene induction. A similar pattern was observed in primary epithelial cells however, the kinetics were slower and depletion was less. In primary airway epithelial cells IkappaBalpha levels were 59.8+/-8.5% of control following 30 min treatment with IL-1beta and in A549 cells 29.1+/-8.5% of control following 15 min IL-1beta treatment. Cytokine-induced IkappaBalpha depletion was associated with NF-kappaB nuclear accumulation and subsequent resynthesis of IkappaBalpha and upregulation of ICAM-1 in both cell types. The antioxidant, NAC (20 mM) had no effect on the kinetics of cytokine-induced IkappaBalpha depletion or NF-kappaB p65 nuclear translocation in either cell type and failed to influence kappaB dependent IkappaBalpha resynthesis. H2O2 treatment alone or in combination with cytokines had no significant effects on IkappaBalpha depletion, NF-kappaB p65 nuclear translocation or ICAM-1 expression in either cell type but did cause significant activation of p38 MAPK. These results suggest that cytokine-induced NF-kappaB activation in cultured human airway epithelial cells does not involve an NAC-sensitive oxidant stress and that H2O2-induced oxidant stress does not result in effective NF-kappaB activation and NF-kappaB mediated gene induction.
Hoare
Ginette S
GS
Department of Cardiothoracic Surgery, National Heart and Lung Institute, Imperial College of Science, Technology and Medicine, Heart Science Centre, Royal Brompton and Harefield NHS Trust, Harefield, Middlesex, UB9 6JH, UK.
Chester
Adrian H
AH
Yacoub
Magdi H
MH
Marczin
Nandor
N
D000111:Acetylcysteine / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis / Q000502:physiology*; D007375:Interleukin-1 / Q000502:physiology; D018613:Microscopy, Confocal; D016328:NF-kappa B / Q000096:biosynthesis / Q000502:physiology*; D016877:Oxidants / Q000378:metabolism*; D018384:Oxidative Stress; D020545:Respiratory Mucosa / Q000502:physiology*; D015533:Transcriptional Activation
D007375:Interleukin-1; D016328:NF-kappa B; D016877:Oxidants; D018799:Intercellular Adhesion Molecule-1; D006861:Hydrogen Peroxide; D000111:Acetylcysteine