RESUMO
The effects of gentamicin on N-acetyl-beta-D-glucosaminidase (NAG) and acid phosphatase (AcP), two lysosomal enzymes present in proximal renal tubule cells, were studied in the PKSV-PCT cell line derived from proximal convoluted tubules from the kidney of a transgenic mouse carrying SV40 large T antigen under the control of the L-type pyruvate kinase gene. Gentamicin (400 micrograms/ml for 72 hr) did not alter cell viability, but significantly reduced cell growth and favored the formation of myeloid bodies. Gentamicin (50 to 800 micrograms/ml for 72 hr) decreased in a dose-dependent manner the cellular NAG in PKSV-PCT cells and stimulated its secretion by 20 to 60%. Chloroquine (50 to 100 microns) and ammonium chloride (NH4Cl, 30mM), two lysosomotropic amines known to stimulate the secretion of lysosomal enzymes in fibroblasts and macrophages, also stimulated secreted NAG in PKSV-PCT cells. However, the effect of chloroquine was less marked in PKSV-PCT cells than in cultured mouse 3T3 fibroblasts. Gentamicin induced lysosomal alkalinization but, in contrast to chloroquine and NH4Cl, the aminoside strongly stimulated the secretion of AcP. The secretion induced by gentamicin was nonpolarized, since the percentage of secreted NAG significantly increased from both the apical and basal sides of PKSV-PCT cells grown on permeable filters. Thus, these data suggest that gentamicin alters the secretion of NAG and AcP by a non-specific pathway and indicate that the PKSV-PCT cell line is a suitable system to examine the cellular action of drugs in kidney proximal tubule cells.