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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...

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Autores principales: Hellermann, Gary R, Nagy, Szilvia B, Kong, Xiaoyuan, Lockey, Richard F, Mohapatra, Shyam S
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2002
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.
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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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