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首页> 外文期刊>The Journal of toxicological sciences >Increased expression of heat shock protein (HSP)72 in a human proximal tubular cell line (HK-2) with gentamicin-induced injury.
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Increased expression of heat shock protein (HSP)72 in a human proximal tubular cell line (HK-2) with gentamicin-induced injury.

机译：在庆大霉素诱导的伤害的人类近端肾小管细胞系（HK-2）中热休克蛋白（HSP）72的表达增加。

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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 activities and results of MDA content were similar to that of the LDH release. The amount of HSP72 positive cells increased at 24 hr after exposure to GM up to 72 hr. HSP72 expression increased significantly from 24 hr, and reached its peak at 72 hr when cells were treated with GM only. Furthermore, the change of the HSP72 gene transcription was similar to the expression of HSP72. These results demonstrated that GM treatment could induce damage to HK-2 cells and that the expression of HSP72 increased when cells were injured by GM.
机译：庆大霉素（GM）已被广泛用作抗生素，其肾毒性已得到公认。然而，目前尚不清楚热激蛋白（HSP）72作为GM处理的近端肾小管细胞中的诱导蛋白的变化。在这项研究中，测量了GM细胞毒性及其对人肾近端肾小管（HK-2）细胞中HSP72表达的影响。 HK-2细胞分别仅与GM和GM加MnCl2一起孵育24小时，48小时，72小时和96小时。细胞毒性通过乳酸脱氢酶（LDH）的释放来确定。通过测量丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性以及存活细胞的能力，研究了N-乙酰基-β-D-氨基葡萄糖苷酶（NAG）的活性以及GM对HK-2细胞氧化的影响。还原基于四唑的化合物（MTT）。通过免疫细胞化学，蛋白质印迹和RT-PCR测量HSP72的表达。将细胞暴露于浓度为100微克/毫升的GM。 24小时后，MTT吸收显着下降，然后逐渐下降直至96小时。与仅使用GM处理细胞时的72小时数据相比，LDH释放时间从24小时增加至72小时，但在96小时时降低。 NAG和SOD活性的结果以及MDA含量的结果均与LDH释放的结果相似。暴露于GM后长达24小时，HSP72阳性细胞的数量增加。当仅用GM处理细胞时，HSP72表达从24小时开始显着增加，并在72小时达到峰值。此外，HSP72基因转录的变化与HSP72的表达相似。这些结果证明了GM处理可以诱导HK-2细胞的损伤，并且当细胞被GM损伤时HSP72的表达增加。

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