title: Gentamicin-induced increases in cytosolic calcium in pig kidney cells (LLC-PK1).
abstract: LLC-PK1 cells, an established epithelial cell line derived from pig kidney, were tested as a model system for assessing the role of calcium in gentamicin-induced nephrotoxicity. Cell viability was evaluated by a vital dye exclusion procedure, and intracellular free calcium [Ca2+]i was measured employing Fura-2 fluorescence. Exposing cell suspensions (10(6)/ml) to concentrations of the drug, which had no apparent effect on viability, produced a rapid and prolonged increase in intracellular [Ca2+]. The perturbation of calcium homeostasis could be blocked by the addition of mepiperphenidol, an inhibitor of the organic cation transport system. We propose that LLC-PK1 cells are an appropriate model to study drug-induced nephrotoxicity. Gentamicin disrupts calcium homeostasis and causes plasma membrane alterations. Since mepiperphenidol blocked the gentamicin-induced Ca2+ increases, the data suggest that aminoglycosides enter the cell via the organic cation transporter.
authors.affiliation: Department of Pharmacology, State University of New York Health Science Center, Syracuse.
authors.forename: P D
authors.initials: PD
authors.lastname: Holohan
authors.forename: P P
authors.initials: PP
authors.lastname: Sokol
authors.forename: C R
authors.initials: CR
authors.lastname: Ross
authors.forename: R
authors.initials: R
authors.lastname: Coulson
authors.forename: M E
authors.initials: ME
authors.lastname: Trimble
authors.forename: D A
authors.initials: DA
authors.lastname: Laska
authors.forename: P D
authors.initials: PD
authors.lastname: Williams
mesh_terms: D000818:Animals; D002118:Calcium / Q000032:analysis*; D002460:Cell Line; D003600:Cytosol / Q000032:analysis*; D004305:Dose-Response Relationship, Drug; D005839:Gentamicins / Q000633:toxicity*; D006706:Homeostasis / Q000187:drug effects; D007668:Kidney / Q000032:analysis / Q000187:drug effects*; D010880:Piperidines / Q000494:pharmacology; D013552:Swine
chemical_list: D005839:Gentamicins; D010880:Piperidines; C010482:mepiperphenidol; D002118:Calcium