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Gentamicin, an aminoglycoside antibiotic, induces apoptosis in the proximal tubule epithelium of rats treated at low, therapeutically relevant doses (El Mouedden et al., Antimicrob. Agents Chemother. 44, 665-675, 2000). Renal cell lines (LLC-PK(1) and MDCK-cells) have been used to further characterize and quantitate this process (electron microscopy; terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling of fragmented DNA [TUNEL]; and DNA size analysis [oligonucleosomal laddering]). Cells were exposed for up to 4 days to gentamicin concentrations of up to 3 mM. Apoptosis developed, almost linearly, with time and drug concentration, and was (i) preventable within the time-frame of the experiments by overexpression of the anti-apoptotic protein Bcl-2, and by co-incubation with cycloheximide (MDKC but not LLC-PK(1) cells); (ii) associated with an increased activity of caspases (MDCK cells; bcl-2 transfectants showed no increase of caspase activities and Z-VAD.fmk afforded full protection). Gentamicin-induced apoptosis also developed to a similar extent in embryonic fibroblasts cultured under the same conditions. In the 3 cell types, apoptosis (measured after 4 days) was directly correlated with cell gentamicin content (apoptotic index [approximately 10 to 18% of TUNEL (+) cells for a content of 20 microg of gentamicin/mg protein; kidney cortex of rats showing apoptosis in proximal tubule epithelium typically contains approximately 10 microg of gentamicin/mg protein). Thus, gentamicin has an intrinsic capability of inducing apoptosis in eucaryotic cells. Development of apoptosis in proximal tubules of kidney cortex in vivo after gentamicin systemic administration is therefore probably related to its capacity to concentrate in this epithelium after systemic administration.

Gentamicin-Induced Apoptosis in Renal Cell Lines and Embryonic Rat Fibroblasts