Eicosapentaenoic acid inhibits TNF-α-induced Lnk expression in human umbilical vein endothelial cells: involvement of the PI3K/Akt pathway

Abstract

n−3 Polyunsaturated fatty acids (PUFAs) exert anti-inflammatory properties by influencing inflammatory cell activation processes. Lnk is an adaptor protein involving endothelial cell (EC) activation because it is induced by tumor necrosis factor-α (TNF-α). This study was conducted to evaluate the role of eicosapentaenoic acid (EPA), an n−3 PUFA, in the regulation of Lnk expression in human umbilical vein endothelial cells (HUVECs). Primary HUVECs were pretreated with EPA for 12 h at various concentrations (0–40 μM) and then exposed for another 12 h in the presence or absence of TNF-α (10 ng/ml). Lnk mRNA and protein were detected using reverse transcriptase polymerase chain reaction, immunoprecipitation and Western blot analysis. Results showed that pretreatment of HUVEC with EPA inhibited TNF-α-induced expression of Lnk in a dose-dependent manner. TNF-α-induced Lnk was also inhibited by a phosphatidylinositol 3-kinase (PI3K) inhibitor, LY294002. Thus, we investigated the role of PI3K/Akt signaling pathway in this process. Phosphorylation of Akt was assessed by Western blot analysis. We found that EPA treatment decreased the amount of activated Akt. These results showed that EPA inhibited TNF-α-induced Lnk expression in HUVECs through the PI3K/Akt pathway. This may be a potential mechanism by which EPA protects ECs under inflammatory conditions.