Document Detail

Alcohol increases the permeability of airway epithelial tight junctions in Beas-2B and NHBE cells.
MedLine Citation:
PMID:  21950588     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: Tight junctions form a continuous belt-like structure between cells and act to regulate paracellular signaling. Protein kinase C (PKC) has been shown to regulate tight junction assembly and disassembly and is activated by alcohol. Previous research has shown that alcohol increases the permeability of tight junctions in lung alveolar cells. However, little is known about alcohol's effect on tight junctions in epithelium of the conducting airways. We hypothesized that long-term alcohol exposure reduces zonula occluden-1 (ZO-1) and claudin-1 localization at the cell membrane and increases permeability through a PKC-dependent mechanism.
METHODS: To test this hypothesis, we exposed normal human bronchial epithelial (NHBE) cells, cells from a human bronchial epithelial transformed cell line (Beas-2B), and Beas-2B expressing a PKCα dominant negative (DN) to alcohol (20, 50, and 100 mM) for up to 48 hours. Immunofluorescence was used to assess changes in ZO-1, claudin-1, claudin-5, and claudin-7 localization. Electric cell-substrate impedance sensing was used to measure the permeability of tight junctions between monolayers of NHBE, Beas-2B, and DN cells.
RESULTS: Alcohol increased tight junction permeability in a concentration-dependent manner and decreased ZO-1, claudin-1, claudin-5, and claudin-7 localization at the cell membrane. To determine a possible signaling mechanism, we measured the activity of PKC isoforms (alpha, delta, epsilon, and zeta). PKCα activity significantly increased in Beas-2B cells from 1 to 6 hours of 100 mM alcohol exposure, while PKCζ activity significantly decreased at 1 hour and increased at 3 hours. Inhibiting PKCα with Gö-6976 prevented the alcohol-induced protein changes in both ZO-1 and claudin-1 at the cell membrane. PKCα DN Beas-2B cells were resistant to alcohol-induced protein alterations.
CONCLUSIONS: These results suggest that alcohol disrupts ZO-1, claudin-1, claudin-5, and claudin-7 through the activation of PKCα, leading to an alcohol-induced "leakiness" in bronchial epithelial cells. Such alcohol-induced airway-leak state likely contributes to the impaired airway host defenses associated with acute and chronic alcohol ingestion.
Samantha M Simet; Todd A Wyatt; Jane DeVasure; Daniel Yanov; Diane Allen-Gipson; Joseph H Sisson
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2011-09-26
Journal Detail:
Title:  Alcoholism, clinical and experimental research     Volume:  36     ISSN:  1530-0277     ISO Abbreviation:  Alcohol. Clin. Exp. Res.     Publication Date:  2012 Mar 
Date Detail:
Created Date:  2012-02-24     Completed Date:  2012-06-22     Revised Date:  2014-09-22    
Medline Journal Info:
Nlm Unique ID:  7707242     Medline TA:  Alcohol Clin Exp Res     Country:  England    
Other Details:
Languages:  eng     Pagination:  432-42     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 by the Research Society on Alcoholism.
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MeSH Terms
Carbazoles / pharmacology
Cell Line
Cell Line, Transformed
Claudins / metabolism
Dose-Response Relationship, Drug
Drug Interactions
Enzyme Inhibitors / pharmacology
Epithelial Cells / drug effects,  metabolism
Ethanol / administration & dosage,  antagonists & inhibitors,  pharmacology*
Isoenzymes / metabolism
Membrane Proteins / metabolism
Permeability / drug effects
Phosphoproteins / metabolism
Protein Kinase C / antagonists & inhibitors,  genetics,  metabolism
Respiratory Mucosa / drug effects*,  metabolism*
Tight Junctions / drug effects*,  metabolism*
Zonula Occludens-1 Protein
Grant Support
5R01AA017993-02/AA/NIAAA NIH HHS; 5R37AA008769-20/AA/NIAAA NIH HHS; R01 AA008769/AA/NIAAA NIH HHS; R01 AA017993/AA/NIAAA NIH HHS; R37 AA008769/AA/NIAAA NIH HHS; R37 AA008769-18/AA/NIAAA NIH HHS; R37 AA008769-19/AA/NIAAA NIH HHS
Reg. No./Substance:
0/CLDN1 protein, human; 0/CLDN5 protein, human; 0/CLDN7 protein, human; 0/Carbazoles; 0/Claudin-1; 0/Claudin-5; 0/Claudins; 0/Enzyme Inhibitors; 0/Isoenzymes; 0/Membrane Proteins; 0/Phosphoproteins; 0/TJP1 protein, human; 0/Zonula Occludens-1 Protein; 136194-77-9/Go 6976; 3K9958V90M/Ethanol; EC Kinase C

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