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Mechanism of cigarette smoke condensate-induced acute inflammatory response in human bronchial epithelial cells
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...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2002
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC150508/ https://www.ncbi.nlm.nih.gov/pubmed/12204101 http://dx.doi.org/10.1186/rr172 |
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author | Hellermann, Gary R Nagy, Szilvia B Kong, Xiaoyuan Lockey, Richard F Mohapatra, Shyam S |
author_facet | Hellermann, Gary R Nagy, Szilvia B Kong, Xiaoyuan Lockey, Richard F Mohapatra, Shyam S |
author_sort | Hellermann, Gary R |
collection | PubMed |
description | 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 μg/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-1β, IL-8 and GM-CSF, as determined by RT-PCR. CSC-induced IL-1β expression was reduced by PD98059, a blocker of mitogen-actived protein kinase (MAPK) kinase (MEK), and by PDTC, a NFκB 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 IκB was also seen after CSC exposure. A549 cells transfected with a luciferase reporter plasmid containing a NFκB-inducible promoter sequence and exposed to CSC (0.4 μg/ml) or TNF-α (50 ng/ml) had an increased reporter activity of approximately 2-fold for CSC and 3.5-fold for TNF-α relative to untreated controls. CONCLUSION: The acute phase response of NHBEs to cigarette smoke involves activation of both MAPK and NFκB. |
format | Text |
id | pubmed-150508 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2002 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-1505082003-03-08 Mechanism of cigarette smoke condensate-induced acute inflammatory response in human bronchial epithelial cells Hellermann, Gary R Nagy, Szilvia B Kong, Xiaoyuan Lockey, Richard F Mohapatra, Shyam S Respir Res Research 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 μg/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-1β, IL-8 and GM-CSF, as determined by RT-PCR. CSC-induced IL-1β expression was reduced by PD98059, a blocker of mitogen-actived protein kinase (MAPK) kinase (MEK), and by PDTC, a NFκB 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 IκB was also seen after CSC exposure. A549 cells transfected with a luciferase reporter plasmid containing a NFκB-inducible promoter sequence and exposed to CSC (0.4 μg/ml) or TNF-α (50 ng/ml) had an increased reporter activity of approximately 2-fold for CSC and 3.5-fold for TNF-α relative to untreated controls. CONCLUSION: The acute phase response of NHBEs to cigarette smoke involves activation of both MAPK and NFκB. BioMed Central 2002 2002-07-10 /pmc/articles/PMC150508/ /pubmed/12204101 http://dx.doi.org/10.1186/rr172 Text en Copyright © 2002 Hellerman et al., licensee BioMed Central Ltd |
spellingShingle | Research Hellermann, Gary R Nagy, Szilvia B Kong, Xiaoyuan Lockey, Richard F Mohapatra, Shyam S Mechanism of cigarette smoke condensate-induced acute inflammatory response in human bronchial epithelial cells |
title | Mechanism of cigarette smoke condensate-induced acute inflammatory response in human bronchial epithelial cells |
title_full | Mechanism of cigarette smoke condensate-induced acute inflammatory response in human bronchial epithelial cells |
title_fullStr | Mechanism of cigarette smoke condensate-induced acute inflammatory response in human bronchial epithelial cells |
title_full_unstemmed | Mechanism of cigarette smoke condensate-induced acute inflammatory response in human bronchial epithelial cells |
title_short | Mechanism of cigarette smoke condensate-induced acute inflammatory response in human bronchial epithelial cells |
title_sort | mechanism of cigarette smoke condensate-induced acute inflammatory response in human bronchial epithelial cells |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC150508/ https://www.ncbi.nlm.nih.gov/pubmed/12204101 http://dx.doi.org/10.1186/rr172 |
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