Gürbay Aylin, Gonthier Brigitte, Barret Luc, Favier Alain, Hincal Filiz
Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical Toxicology, 06100 Ankara, Turkey.
Toxicology. 2007 Jan 5;229(1-2):54-61. doi: 10.1016/j.tox.2006.09.016. Epub 2006 Oct 1.
The aim of this study was to investigate the possible cytotoxic and oxidative stress inducing effects of ciprofloxacin (CPFX) on primary cultures of rat astrocytes. The cultured cells were incubated with various concentrations of CPFX (0.5-300mg/l), and cytotoxicity was determined by neutral red (NR) and MTT assays. Survival profile of cells was biphasic in NR assay: CPFX did not cause any alteration at any concentration for 7h, whereas < or =50mg/l concentrations induced significant cell proliferation in incubation periods of 24, 48, 72, and 96h. However, cell proliferation gradually decreased at higher concentrations, and 200 and 300mg/l of CPFX exposure was found to be significantly (p<0.05) cytotoxic at all time periods. With MTT assay, no alteration was noted for incubation period of 7h, as observed with NR assay. But, cell viability decreased with approximately > or =50mg/l CPFX exposure in all other time periods. Cell proliferation was only seen in 24h of incubation with 0.5 and 5mg/l CPFX. Vitamin E pretreatment of cell cultures were found to be providing complete protection against cytotoxicity of 300mg/l CPFX in 96h incubation when measured with both NR and MTT assays. The SOD pretreatment was partially protective with NR assay, but no protection was noted when measured with MTT. A significant enhancement of lipid peroxidation was observed with the cytotoxic concentration of the drug, but total glutathione content and catalase activity of cells did not change. The data obtained in this study suggest that, in accordance with our previous results with fibroblast cells, CPFX-induced cytotoxicity is related to oxidative stress. And the biphasic effect of CPFX possibly resulted from the complex dose-dependent relationships between reactive oxygen species, cell proliferation, and cell viability.
本研究的目的是调查环丙沙星（CPFX）对大鼠星形胶质细胞原代培养物可能产生的细胞毒性和氧化应激诱导作用。将培养的细胞与不同浓度的CPFX（0.5 - 300mg/l）孵育，并用中性红（NR）和MTT法测定细胞毒性。在NR试验中，细胞存活情况呈双相性：CPFX在任何浓度下7小时内均未引起任何改变，而≤50mg/l的浓度在24、48、72和96小时的孵育期内诱导细胞显著增殖。然而，在较高浓度下细胞增殖逐渐降低，并且发现200和300mg/l的CPFX暴露在所有时间段均具有显著（p<0.05）细胞毒性。与NR试验一样，MTT试验在7小时的孵育期内未观察到任何改变。但是，在所有其他时间段，当CPFX暴露浓度≥50mg/l时细胞活力下降。仅在0.5和5mg/l CPFX孵育24小时时观察到细胞增殖。当用NR和MTT试验测定时，发现细胞培养物用维生素E预处理可在96小时孵育中完全保护细胞免受300mg/l CPFX的细胞毒性。SOD预处理在NR试验中具有部分保护作用，但用MTT测定时未观察到保护作用。在药物的细胞毒性浓度下观察到脂质过氧化显著增强，但细胞的总谷胱甘肽含量和过氧化氢酶活性未改变。本研究获得的数据表明，与我们先前对成纤维细胞的研究结果一致，CPFX诱导的细胞毒性与氧化应激有关。CPFX的双相效应可能源于活性氧、细胞增殖和细胞活力之间复杂的剂量依赖关系。
