Premium
Alcohol‐induced decreases in brain endothelial barrier integrity rescued by sphingosine‐1‐phosphate or 8‐pCPT‐2'‐O‐Me‐cAMP
Author(s) -
Alves Natascha,
Adderley Shaquria,
Yuan Sarah,
Breslin Jerome
Publication year - 2015
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.29.1_supplement.791.8
Subject(s) - tight junction , sphingosine 1 phosphate , blood–brain barrier , barrier function , chemistry , sphingosine , microbiology and biotechnology , intracellular , sphingosine kinase 1 , evans blue , permeability (electromagnetism) , endocrinology , medicine , biochemistry , biology , central nervous system , membrane , receptor
Alcohol has previously been shown to increase blood‐brain‐barrier (BBB) permeability. We hypothesized that alcohol increases BBB permeability by disrupting the junctional proteins, VE‐cadherin and claudin‐5, and that barrier enhancers such as sphingosine‐1‐phosphate (S1P) and 8‐pCPT‐2'‐O‐Me‐cAMP (CPT) can restore barrier function. We also hypothesized that p38 MAP kinase mediates alcohol‐induced BBB dysfunction. Primary human brain microvascular endothelial cells (HBMEC) were used to model the BBB in vitro . Transendothelial electrical resistance (TER) served as an index of barrier function. Protein localization was determined by immunofluorescence microscopy. Alcohol (50‐100 mM) rapidly and significantly decreased TER. S1P (1 μM) or CPT (100 μM) significantly enhanced barrier function. Application of S1P or CPT after alcohol (75 mM) significantly decreased the time for TER to recover to baseline levels, compared to alcohol alone. Pretreating HMBEC with the p38 MAP kinase inhibitor SB208530 (20 μM) did not affect the alcohol‐induced decrease in TER. Immunofluorescence data showed localization of VE‐cadherin and claudin‐5 at intercellular junctions, which was disrupted by alcohol treatment. Treatment with S1P after alcohol restored junctional integrity of VE‐cadherin and claudin‐5. Our results demonstrate that alcohol weakens brain endothelial barrier integrity, and that S1P and CPT can enhance barrier function. In addition, our data shows that S1P can restore the integrity of intercellular junctions. These data suggest S1P or activation of Epac1 may be useful to ameliorate BBB dysfunction. Supported by NIH HL098215, HL070752, and GM097270.