Gentamicin nephrotoxicity--a comparison of in vitro findings with in vivo experiments in equines.
The aminoglycoside gentamicin is often used in equine practice. Despite its clinical use, concerns remain regarding the potential toxic side-effects, such as nephrotoxicity, in equine patients, particularly after repeated dosing. The aim of the study was to investigate first in vitro the mechanisms contributing to the renal toxicity of gentamicin and to identify sensitive biomarkers indicating proximal tubule damage. To this end, the kidney-derived cell lines LLC-PKI and MDCK were treated with gentamicin at different concentrations. Toxicity was assessed by measuring the release of gamma-glutamyl transferase (GGT), and the production of reactive oxygen species (ROS). Cell viability was measured using Alamar blue (AB) and Neutral red (NR) cytotoxicity assays. Gentamicin exerted a dose-dependent toxicity. Primarily, loss of brush border membrane integrity, indicated by GGT leakage, and an increased ROS production were observed. As GGT was found to be a sensitive marker for gentamicin-induced renal cell injury, in the subsequent in vivo experiments, in which ponies were given gentamicin (3.0 mg/kg bw three times daily and 4.5 mg/kg bw twice daily) for five consecutive days, plasma levels and the urinary excretion of GGT and creatinine were measured and the GGT:creatinine ratio was calculated. Elevated GGT levels in urine following gentamicin therapy were observed, but this enzyme leakage was transient and returned to baseline values after cessation of therapy. It could thus be concluded that even a conservative dose regimen of gentamicin did not result in significant renal toxicity in healthy ponies.
van der Harst
M R
MR
Faculty of Veterinary Medicine, Department of Equine Sciences, Utrecht University, Yalelaan 12, 3584 CM Utrecht, The Netherlands. m.r.vanderHarst@vet.uu.nl
Bull
S
S
Laffont
C M
CM
Klein
W R
WR
D000818:Animals; D000900:Anti-Bacterial Agents / Q000493:pharmacokinetics / Q000633:toxicity*; D015415:Biomarkers / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D003404:Creatinine / Q000378:metabolism; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D016231:Fluorescence Polarization Immunoassay / Q000662:veterinary; D005839:Gentamicins / Q000493:pharmacokinetics / Q000633:toxicity*; D006736:Horses; D007686:Kidney Tubules, Distal / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D018374:LLC-PK1 Cells; D008297:Male; D009499:Neutral Red; D010078:Oxazines; D017382:Reactive Oxygen Species / Q000378:metabolism; D013552:Swine; D014966:Xanthenes; D005723:gamma-Glutamyltransferase / Q000378:metabolism
D000900:Anti-Bacterial Agents; D015415:Biomarkers; D005839:Gentamicins; D010078:Oxazines; D017382:Reactive Oxygen Species; D014966:Xanthenes; C005843:resazurin; D009499:Neutral Red; D003404:Creatinine; D005723:gamma-Glutamyltransferase