Investigations of oxidative stress, antioxidant response, and protein binding in chlorpyrifos exposed rat neuronal PC12 cells
Investigations of oxidative stress, antioxidant response, and protein binding in chlorpyrifos exposed rat neuronal PC12 cells
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DOI：
10.1080/15376510701389530
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发表时间：
2008-01-01
影响因子：
3.2
通讯作者：
Gollapudi, Bhaskar B.
中科院分区：
文献类型：
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作者：
Geter, David R.;Kan, H. Lynn;Gollapudi, Bhaskar B.
毒死蜱是一种广泛使用的有机磷杀虫剂。除了已知的胆碱酯酶抑制特性外，活性氧（ROS）的产生也被认为是一种可能的毒性机制。为了研究CPF产生的ROS，将大鼠神经元PC 12细胞暴露于浓度为0至5000 μ g/mL的CPF，其在Krebs缓冲培养基（KRH）、KRH + 4%牛血清白蛋白（BSA）和KRH + 25 μ M抗氧化剂Trolox中0至5小时。百草枯作为活性氧的阳性对照。用荧光探针2，7-二氯二氢荧光素和MTS法分别测定ROS和细胞毒性。还进行了CPF-白蛋白结合检查。CPF对PC 12细胞没有强烈的细胞毒性，在5000 μ g/ml时仅引起轻度细胞毒性。在KRH培养基中，在500和1000 μ g/mL CPF下，在4和5 h观察到CPF产生的ROS，在5000 μ g/mL CPF下，在1至5 h观察到CPF产生的ROS。在KRH +4%BSA中，仅在5000 μ g/mL CPF中的5 h时观察到ROS。Trolox显着降低CPF和百草枯诱导的ROS。在4%BSA中，1、10和100 μ g/mL CPF的计算CPF-白蛋白结合率分别为96%、75%和15%。这些数据显示CPF >= 500 μ g/mL在PC 12细胞中诱导ROS，但添加抗氧化剂Trolox和4%BSA显著降低ROS水平。
Chlorpyrifos (CPF) is a widely used organophosphate insecticide. In addition to its known properties of cholinesterase inhibition, the production of reactive oxygen species (ROS) has been suggested as a possible toxic mechanism. To investigate CPF-generated ROS, rat neuronal PC12 cells were exposed to CPF concentrations of 0 to 5000 mu g/mL in Krebs buffered media (KRH), KRH + 4% bovine serum albumin (BSA), and KRH + 25 mu M of the antioxidant Trolox for 0 to 5 h. Paraquat served as a positive control for ROS. The fluorescent probe 2,7-dichlorodihydro-fluorescein and the MTS assay were used to measure ROS and cytotoxicity, respectively. Examinations into CPF-albumin binding were also conducted. CPF was not strongly cytotoxic to PC12 cells, causing only mild cytotoxicity at 5000 mu g/ml. In KRH media, CPF-generated ROS was observed at 4 and 5 h at 500 and 1000 mu g/mL, and at 1 to 5 h at 5000 mu g/mL CPF. In KRH + 4% BSA, ROS was seen only at 5 h in 5000 mu g/mL CPF. Trolox significantly reduced CPF- and paraquat-induced ROS. Calculated CPF-albumin binding at 1, 10, and 100 g/mL CPF in 4% BSA was 96%, 75%, and 15%. These data show CPF >= 500 g/mL induced ROS in PC12 cells, but the addition of the antioxidant Trolox and 4% BSA dramatically reduced ROS levels.