Stabilizing effects of eicosapentaenoic acid on Kv1.5 channel protein expressed in mammalian cells

• Published in: European journal of pharmacology Vol. 604; no. 1; pp. 93 - 102
• Main Authors: Koshida, Shunya, Kurata, Yasutaka, Notsu, Tomomi, Hirota, Yutaka, Kuang, Ting Y., Li, Peili, Bahrudin, Udin, Harada, Shingo, Miake, Junichiro, Yamamoto, Yasutaka, Hoshikawa, Yoshiko, Igawa, Osamu, Higaki, Katsumi, Soma, Masaaki, Yoshida, Akio, Ninomiya, Haruaki, Shiota, Goshi, Shirayoshi, Yasuaki, Hisatome, Ichiro
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 14-02-2009
• Subjects: Action Potentials - drug effects; Animals; Blotting, Western; Cercopithecus aethiops; COS Cells; DHA; Docosahexaenoic Acids - pharmacology; Dose-Response Relationship, Drug; Eicosapentaenoic Acid - analogs & derivatives; Eicosapentaenoic Acid - pharmacology; EPA; Fibroblasts - drug effects; Fibroblasts - enzymology; Fibroblasts - metabolism; Immunoprecipitation; Kv1.5; Kv1.5 Potassium Channel - biosynthesis; Oligopeptides; Patch-Clamp Techniques; Peptides - metabolism; Proteasome Endopeptidase Complex - metabolism; Protein Stability; Rats; Rats, Inbred WKY; Recombinant Fusion Proteins - metabolism; Thioctic Acid - analogs & derivatives; Thioctic Acid - pharmacology; Transfection; Ultra-rapid delayed-rectifier K + channel current; Kv1.5; DHA; Ultra-rapid delayed-rectifier K + channel current; EPA
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/j.ejphar.2008.12.016

We investigated the effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on the stability of Kv1.5 channel protein. The expression and function of Kv1.5 (Kv1.5-FLAG) in transfected African green monkey kidney fibroblast cells as well as rat atrium were estimated by immunoblotting, immunoprecipitation, immunofluorescence and patch-clamp techniques. Both EPA and DHA immediately blocked Kv1.5 channel current in a dose-dependent manner, accompanied by reduction of their phosphorylation. Chronic treatment (for 12 h) with EPA at lower concentrations (0.3–10 μM) increased the level of Kv1.5-FLAG protein as well as Kv1.5 channel current without changes in its gating kinetics, prolonging its half-life; in contrast, both EPA and DHA at higher concentrations (30–100 μM) decreased the expression of Kv1.5-FLAG. EPA at the higher concentrations also decreased mRNA of Kv1.5 and synapse-associated protein 97 expression. EPA at the lower concentrations increased Kv1.5 expression in the endoplasmic reticulum, Golgi apparatus and cell membrane. EPA-induced increase of Kv1.5 channel expression and current was abolished by pretreatment with the protein transport inhibitor brefeldin A or colchicines, and by the Kv1.5 channel blocker 4-aminopyridine. Oral administration of EPA (30 mg/kg) increased the level of endogenous Kv1.5 in rat atria. These results indicate that chronic treatment with EPA at lower concentrations stabilizes Kv1.5 channel protein in the endoplasmic reticulum and Golgi apparatus thereby enhancing the Kv1.5 channel current on the cell membrane.