title: Depolarization and decreased surface expression of K+ channels contribute to NSAID-inhibition of intestinal restitution.
abstract: Non-steroidal anti-inflammatory drugs (NSAIDs) contribute to gastrointestinal ulcer formation by inhibiting epithelial cell migration and mucosal restitution; however, the drug-affected signaling pathways are poorly defined. We investigated whether NSAID inhibition of intestinal epithelial migration is associated with depletion of intracellular polyamines, depolarization of membrane potential (E(m)) and altered surface expression of K(+) channels. Epithelial cell migration in response to the wounding of confluent IEC-6 and IEC-Cdx2 monolayers was reduced by indomethacin (100 microM), phenylbutazone (100 microM) and NS-398 (100 microM) but not by SC-560 (1 microM). NSAID-inhibition of intestinal cell migration was not associated with depletion of intracellular polyamines. Treatment of IEC-6 and IEC-Cdx2 cells with indomethacin, phenylbutazone and NS-398 induced significant depolarization of E(m), whereas treatment with SC-560 had no effect on E(m). The E(m) of IEC-Cdx2 cells was: -38.5+/-1.8 mV under control conditions; -35.9+/-1.6 mV after treatment with SC-560; -18.8+/-1.2 mV after treatment with indomethacin; and -23.7+/-1.4 mV after treatment with NS-398. Whereas SC-560 had no significant effects on the total cellular expression of K(v)1.4 channel protein, indomethacin and NS-398 decreased not only the total cellular expression of K(v)1.4, but also the cell surface expression of both K(v)1.4 and K(v)1.6 channel subunits in IEC-Cdx2. Both K(v)1.4 and K(v)1.6 channel proteins were immunoprecipitated by K(v)1.4 antibody from IEC-Cdx2 lysates, indicating that these subunits co-assemble to form heteromeric K(v) channels. These results suggest that NSAID inhibition of epithelial cell migration is independent of polyamine-depletion, and is associated with depolarization of E(m) and decreased surface expression of heteromeric K(v)1 channels.
authors.affiliation: Department of Anatomy and Physiology, Kansas State University, Manhattan, KS 66506, United States.
authors.forename: L C
authors.initials: LC
authors.lastname: Freeman
authors.forename: D F
authors.initials: DF
authors.lastname: Narvaez
authors.forename: A
authors.initials: A
authors.lastname: McCoy
authors.forename: F B
authors.initials: FB
authors.lastname: von Stein
authors.forename: S
authors.initials: S
authors.lastname: Young
authors.forename: K
authors.initials: K
authors.lastname: Silver
authors.forename: S
authors.initials: S
authors.lastname: Ganta
authors.forename: D
authors.initials: D
authors.lastname: Koch
authors.forename: R
authors.initials: R
authors.lastname: Hunter
authors.forename: R F
authors.initials: RF
authors.lastname: Gilmour
authors.forename: J D
authors.initials: JD
authors.lastname: Lillich
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000633:toxicity*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007213:Indomethacin / Q000633:toxicity; D007413:Intestinal Mucosa / Q000187:drug effects* / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects*; D009578:Nitrobenzenes / Q000633:toxicity; D018408:Patch-Clamp Techniques; D010653:Phenylbutazone / Q000633:toxicity; D011073:Polyamines / Q000032:analysis / Q000378:metabolism; D051637:Shaker Superfamily of Potassium Channels / Q000187:drug effects* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D021241:Spectrometry, Mass, Electrospray Ionization; D013449:Sulfonamides / Q000633:toxicity; D014945:Wound Healing / Q000187:drug effects* / Q000502:physiology
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D009578:Nitrobenzenes; D011073:Polyamines; D051637:Shaker Superfamily of Potassium Channels; D013449:Sulfonamides; C080955:N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; D010653:Phenylbutazone; D007213:Indomethacin