Pretreatment with eicosapentaenoic acid prevented hypoxia/reoxygenation-induced abnormality in endothelial gap junctional intercellular communication through inhibiting the tyrosine kinase activity.
Eicosapentaenoic acid (EPA) may protect against atherosclerotic disease, and modulation of endothelium function is one possible mechanism. Hypoxia/reoxygenation (H/R) is a potential risk factor for the pathogenesis of atherosclerosis, and it causes endothelial dysfunction. To evaluate whether EPA may improve the endothelial dysfunction under the condition of H/R, we examined endothelial gap junctional intercellular communication (GJIC), which is said to be important for the endothelium to maintain its normal function. The results indicate that H/R induced a temporal reduction in GJIC after 2 h of reoxygenation in cultured human umbilical vein endothelial cells (HUVEC). This reduction in GJIC was not observed in cells pretreated with 3 microg/ml EPA for 2 days. The results of immunofluorescence show that 2 h reoxygenation caused an increased production of tyrosine-phosphorylated proteins, which was inhibited by EPA pretreatment. Immunoprecipitation demonstrated that tyrosine residues of connexin 43 (Cx43), an important gap junctional protein in HUVEC, were phosphorylated by H/R. However, pretreatment with EPA significantly suppressed this increased phosphorylation. The protective effect of EPA on the reduction in GJIC was also observed in cells treated with 1.5 mM vanadate, a tyrosine phosphatase inhibitor. These data suggest that EPA may ameliorate the H/R-induced GJIC abnormality via inhibition of the tyrosine kinase activation.
Zhang
Y W
YW
Department of Physiological Chemistry, Graduate School, Tokyo Medical and Dental University, Japan.
Morita
I
I
Yao
X S
XS
Murota
S
S
D015687:Cell Hypoxia / Q000187:drug effects*; D002478:Cells, Cultured; D018031:Connexin 43 / Q000378:metabolism; D015118:Eicosapentaenoic Acid / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000648:ultrastructure*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D017629:Gap Junctions / Q000187:drug effects*; D006801:Humans; D007163:Immunosorbent Techniques; D007700:Kinetics; D010100:Oxygen / Q000008:administration & dosage*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D014471:Umbilical Veins
D018031:Connexin 43; D004791:Enzyme Inhibitors; D019000:Phosphotyrosine; D015118:Eicosapentaenoic Acid; D011505:Protein-Tyrosine Kinases; D010100:Oxygen