Optimisation of cell-based assays for medium throughput screening of oxidative stress
Optimisation of cell-based assays for medium throughput screening of oxidative stress
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DOI：
10.1016/s0887-2333(03)00005-5
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发表时间：
2003-04-01
影响因子：
3.2
通讯作者：
Carmichael, N
中科院分区：
文献类型：
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作者：
Lautraite, S;Bigot-Lasserre, D;Carmichael, N
在筛选化学品的潜在毒性特性时，识别和开发适合中等通量筛选要求的基于细胞的测定法是重要的。我们在此描述了基于荧光计和荧光计的快速微孔板测定法，其允许通过测量三种标志物来评价肝细胞培养物中的氧化应激：通过DCFH-DA探针评估氢过氧化物的产生，通过测定还原型谷胱甘肽和谷胱甘肽过氧化物酶活性测定细胞抗氧化状态，通过用罗丹明123评价线粒体跨膜电位测定细胞毒性和线粒体损伤荧光染料。这里描述的测定是快速、简单和廉价的，这在建立筛选测定时是期望的。该系统应可用于在农用化学品或药物的预开发阶段期间选择候选化合物。它也应该是在体内观察到的潜在毒性机制的回顾性和解释性研究的兴趣。(C)2003爱思唯尔科技有限公司版权所有。
Identification and development of cell-based assays adapted to medium throughput screening requirements is important when screening chemicals for their potential toxic properties. We describe here rapid fluorometer-based and spectrophotometer-based microplate assays which allow for the evaluation of oxidative stress in hepatocyte cell cultures by measurement of three markers: production of hydroperoxide assessed by the DCFH-DA probe, cellular antioxidant status by measurement of reduced glutathione and glutathione peroxidase activity and cytotoxicity and mitochondrial damage by evaluation of the mitochondrial transmembrane potential with rhodamine 123 fluorescent dye. The assays described here are rapid, simple and inexpensive, which are desirable when setting up screening assays. This system should be useful in selecting candidate compounds during the pre-development phase of agrochemicals or pharmaceuticals. It should also be of interest for retrospective and explanatory studies of mechanisms underlying toxicity observed in vivo. (C) 2003 Elsevier Science Ltd. All rights reserved.