Gentamicin-induced increases in cytosolic calcium in pig kidney cells (LLC-PK1).
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.
Holohan
P D
PD
Department of Pharmacology, State University of New York Health Science Center, Syracuse.
Sokol
P P
PP
Ross
C R
CR
Coulson
R
R
Trimble
M E
ME
Laska
D A
DA
Williams
P D
PD
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
D005839:Gentamicins; D010880:Piperidines; C010482:mepiperphenidol; D002118:Calcium