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Alcohol intoxication increases microvascular permeability through inactivation of Rac1 and disruption of VE‐cadherin organization
Author(s) -
Doggett Travis,
Breslin Jerome
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.896.6
Subject(s) - chemistry , vascular permeability , ve cadherin , rac1 , cadherin , microcirculation , umbilical vein , medicine , microbiology and biotechnology , cell , biochemistry , biology , in vitro , signal transduction
We tested the hypothesis that alcohol increases endothelial permeability by inactivating Rac1 and disrupting VE‐cadherin localization. Conscious rats received 2.5 g/kg alcohol or isovolumic water via pre‐implanted gastric catheters. After 30 min, they were anesthetized, prepped for intravital microcopy of the mesenteric microcirculation, and the integrated optical intensity (IOI) of extravasated FITC‐albumin was determined. Transendothelial electrical resistance (TER) of human umbilical vein endothelial cell monolayers also served as an index of barrier function. VE‐cadherin localization was determined by immunofluorescence labeling or in live cells expressing VE‐cadherin‐GFP. Rac1‐GTP was determined by ELISA. The results show that alcohol increases microvascular leak (change in IOI 33.0 ± 16.1 for alcohol vs. 2.86 ± 0.7 for water, p<0.05), and decreases TER when applied at >;20 mM. Alcohol also disrupted VE‐cadherin continuity and significantly decreased Rac1‐GTP (53% ± 5 of control, p<0.01). Addition of 100 μM 8‐CPT‐2′‐O‐Me‐cAMP, an indirect activator of Rac1, at 5 or 10 min post alcohol caused a significantly faster return to baseline TER (respectively 14.9 ± 0.5 and 19.5 ± 0.4 min vs. 59.1 ± 1.5 without activator, p<0.01). The data suggest that alcohol increases endothelial permeability by inactivating Rac1 and disrupting VE‐cadherin at junctions. Supported by NIH R21AA020049 and T32AA007577.