Journal
RESPIRATORY RESEARCH
Volume 3, Issue 1, Pages -Publisher
BMC
DOI: 10.1186/rr172
Keywords
bronchial; cigarette; MAPK; NF-kappaB; signal transduction
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Background: To demonstrate the involvement of tobacco smoking in the pathophysiology of lung disease, the responses of pulmonary epithelial cells to cigarette smoke condensate (CSC) - the particulate fraction of tobacco smoke - were examined. Methods: The human alveolar epithelial cell line A549 and normal human bronchial epithelial cells (NHBEs) were exposed to 0.4 mug/ml CSC, a concentration that resulted in >90% cell survival and <5% apoptosis. Changes in gene expression and signaling responses were determined by RT-PCR, western blotting and immunocytofluorescence. Results: NHBEs exposed to CSC showed increased expression of the inflammatory mediators sICAM-1, IL-1, IL-8 and GM-CSF, as determined by RT-PCR. CSC-induced IL-1 beta expression was reduced by PD98059, a blocker of mitogen-actived protein kinase (MAPK) kinase (MEK), and by PDTC, a NF kappa B inhibitor. Analysis of intracellular signaling pathways, using antibodies specific for phosphorylated MAPKs (extracellular signal-regulated kinase [ERK]-1/2), demonstrated an increased level of phosphorylated ERK1/2 with increasing CSC concentration. Nuclear localization of phosphorylated ERK1/2 was seen within 30 min of CSC exposure and was inhibited by PD98059. Increased phosphorylation and nuclear translocation of I kappa B was also seen after CSC exposure. A549 cells transfected with a luciferase reporter plasmid containing a NF kappa B-inducible promoter sequence and exposed to CSC (0.4 mu g/ml) or TNF-alpha (50 ng/ml) had an increased reporter activity of approximately 2-fold for CSC and 3.5-fold for TNF-alpha relative to untreated controls. Conclusion: The acute phase response of NHBEs to cigarette smoke involves activation of both MAPK and NF kappa B.
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