Cytotoxic potency of H2O2 in cell cultures: Impact of cell concentration and exposure time
Cytotoxic potency of H2O2 in cell cultures: Impact of cell concentration and exposure time
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DOI：
10.1016/j.freeradbiomed.2010.07.015
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发表时间：
2010-11-01
影响因子：
7.4
通讯作者：
Seibert, Hasso
中科院分区：
文献类型：
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作者：
Guelden, Michael;Jess, Anne;Seibert, Hasso
在这项研究中，我们使用C6胶质瘤细胞，详细研究了暴露时间和细胞浓度如何影响过氧化氢在体外的细胞毒效力。随着孵育时间从1小时增加到24小时，中位细胞毒性浓度（EC 50）从500 μ M降低到30 μ M。证明24小时足以确定H2 O2的初始细胞毒性浓度。初始EC 50值与细胞浓度线性相关。得出细胞浓度非依赖性细胞毒性细胞剂量（ED 50）中位数为430 nmol/mg细胞蛋白或860 nmol/10（7）个细胞。中位细胞毒性H2 O2浓度从培养基中完全消除，其速率与H2 O2和细胞浓度成比例。相反，EC 50值的浓度与时间曲线（AUC）下的相应面积是独立的细胞浓度和达1800亩M ×分钟。随着细胞浓度的降低，H2 O2消除减速，因此，暴露于H2 O2作为一个团注应用接近连续暴露于一个稳定的H2 O2浓度。两者合计，我们的结果表明，H2 O2的细胞毒性效力的特点是由AUC，这取决于其初始浓度，细胞消除H2 O2的能力，和细胞浓度。我们建议通过初始毒性细胞剂量（例如，nmol H2 O2/mg细胞蛋白或nmol H2 O2/10（7）个细胞），特别是用于比较目的。(C)2010年爱思唯尔公司All rights reserved.
Using C6 glioma cells in this study we investigated in detail how exposure time and cell concentration affect the cytotoxic potency of H2O2 in vitro. Median cytotoxic concentrations (EC50) decreased from 500 to 30 mu M with increasing incubation time from 1 to 24 h. Twenty-four hours proved to be sufficient to determine incipient cytotoxic concentrations of H2O2. The incipient EC50 values were linearly related to the cell concentration. A cell concentration-independent median cytotoxic cell dose (ED50) of 430 nmol/mg cell protein or 860 nmol/10(7) cells was derived. Median cytotoxic H2O2 concentrations were completely eliminated from the culture medium at a rate proportional to both the H2O2 and the cell concentrations. In contrast to EC50 values the corresponding areas under the concentration versus time curve (AUC) were independent of the cell concentration and amounted to 1800 mu M x min. With decreasing cell concentration the H2O2 elimination decelerates and, thus, exposure to H2O2 applied as a bolus approaches a continuous exposure to a steady H2O2 concentration. Taken together, our results indicate that the cytotoxic potency of H2O2 administered to cultured cells as a bolus is characterized by the AUC, which depends on its initial concentration, the ability of the cells to eliminate H2O2, and the cell concentration. We recommend expressing the toxic potency of H2O2 in vitro by the incipient toxic cell dose (e.g., nmol H2O2/mg cell protein or nmol H2O2/10(7) cells), in particular for comparative purposes. (C) 2010 Elsevier Inc. All rights reserved.