Abstract
Gentamicin (GM) has been widely used as an antibiotic and its nephrotoxicity has been recognized. However, the alternation of heat shock protein (HSP) 72 as an inductive protein in proximal tubular cells treated with GM is still unclear. In this study, GM cytotoxicity and its effect on the expression of HSP72 in human kidney proximal tubular (HK-2) cells were measured. HK-2 cells were incubated for 24 hr, 48 hr, 72 hr, and 96 hr with GM only and GM plus MnCl2, respectively. Cytotoxicity was determined by the release of lactate dehydrogenase (LDH). Activity of N-acetyl-beta-D-glucosaminidase (NAG) and effects of GM on oxidation in HK-2 cells were investigated by measurements of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity, and the ability of viable cells to reduce a tetrazolium-based compound (MTT). The expression of HSP72 was measured by immunocytochemistry, Western blotting and RT-PCR. Cells were exposed to GM at a concentration of 100 microg/ml. After 24 hr MTT uptake decreased significantly and then gradually until 96 hr. LDH release increased time-dependently from 24 hr to 72 hr, but decreased at 96 hr compared with the data at 72 hr when cells were treated with GM only. Both results of NAG and SOD ...Continue Reading
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