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B Activation in Human Bronchial Epithelial Cells In Vitro: Importance in Cytokine Transcription1
Departments of
*
Laboratory Medicine and
Medicine and Physical Therapy, University of Tokyo School of Medicine, Tokyo, Japan;
First Department of Internal Medicine, Showa University School of Medicine, Tokyo, Japan;
§
National Institute of Infectious Disease, Tokyo, Japan;
¶
Department of Molecular Preventive Medicine, University of Tokyo Graduate School of Medicine, Tokyo, Japan; and
||
Fourth Department of Internal Medicine, Nippon Medical School, Tokyo, Japan
Fine particles derived from diesel engines (diesel exhaust
particles, DEP) have attracted attention, since their density in
industrial countries seems related to the increased prevalence of
pulmonary diseases. Previous studies have suggested that DEP have a
potential to directly activate airway epithelial cells to produce and
release inflammatory cytokines and mediators, and thus facilitate
inflammatory responses in the lung. To elucidate the molecular
mechanisms of their action, we studied here IL-8 gene expression, one
of the important cytokines in inflammatory responses, by Northern blot
analysis and run-on transcription assay. Suspended DEP (1–50 µg/ml)
increased the steady state levels of IL-8 mRNA, which was suggested to
be largely due to increased transcriptional rates. Electrophoretic
mobility shift assay demonstrated that DEP induced increased binding to
the specific motif of NF-
B, but not of transcription factor AP-1.
The luciferase reporter gene assay using wild-type and mutated
NF-B-binding sequences showed that DEP-induced NF-B activation
was involved in IL-8 transcription. Finally, both
N-acetylcysteine and pyrrolidine dithiocarbamate
attenuated the action of DEP on IL-8 mRNA expression, suggesting that
oxidant-mediated pathway might be involved in its processes. These
results suggested that DEP activate NF-B, which might be an
important mechanism of its potential to increase the expression of
inflammatory cytokines in vitro.
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