ENGLISH ABSTRACT
JOURNAL ARTICLE
RESEARCH SUPPORT, NON-U.S. GOV'T
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[Effects of chronic ethanol ingestion on intercellular junction proteins and barrier function of airway epithelium in the endotoxemic rat].

OBJECTIVE: To investigate the effects of chronic ethanol ingestion and lipopolysaccharide (LPS) on airway epithelium barrier function and the expression of tight junction protein occludin and adherens junction protein E-cadherin.

METHODS: Forty Sprague-Dawley (SD) rats were assigned to control, ethanol, LPS and ethanol+LPS groups. The bronchoalveolar epithelial permeability was assessed by the bronchoalveolar lavage fluid (BALF): serum fluorescein isothiocyanate (FITC)-dextran (FD(4)) fluorescence ratio. Protein localization and expression of occludin and E-cadherin in airway epithelium of rats lung were examined by immunofluorescence. The protein, messenger RNA expression of occludin and E-cadherin in the lung were examined by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTS: The bronchoalveolar epithelial permeability in the ethanol group and LPS group were approximately 2-fold higher than those in the control group (both P<0.05). The permeability in the ethanol+LPS group was increased further (P<0.01). Occludin and E-cadherin appeared as a continuous and uniform expression in the airway epithelium of control rats. In ethanol group and LPS group partial breakdown of membrane staining and decreased cytoplasm staining of occludin and E-cadherin were seen in the airway epithelium. In ethanol plus LPS group, there was breakdown of epithelial membrane and obvious disappearance of cytoplasm staining pattern of occludin and E-cadherin. Western blotting and RT-PCR showed that both in ethanol and LPS groups protein levels of occludin and E-cadherin were depressed in lung. Treatment with ethanol together with LPS further depressed the protein and mRNA levels of occludin and E-cadherin in lung. RT-PCR showed that messenger RNA levels of occludin and E-cadherin in ethanol group and LPS group were lower than those in the control group (both P< 0.05). Treatment with ethanol plus LPS had the greatest depressive effect on these messenger RNA levels (ethanol+LPS group vs. the other groups, all P<0.01).

CONCLUSION: These data suggest that chronic ethanol ingestion impairs the airway epithelial barrier function through down-regulation the messenger RNA expression and disruption of membrane localization of occludin and E-cadherin. Those deterioration of occludin and E-cadherin predisposes to acute lung injury induced by LPS.

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