Cytotoxicity and genotoxicity of urban particulate matter in mammalian cells.
Cytotoxicity and genotoxicity of urban particulate matter in mammalian cells.
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DOI：
10.1093/mutage/gev025
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发表时间：
2015-09
期刊：
影响因子：
2.7
通讯作者：
Povey AC
中科院分区：
文献类型：
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作者：
Dumax-Vorzet AF;Tate M;Walmsley R;Elder RH;Povey AC
环境空气颗粒物(PM)相关的活性氧物种(ROS)与细胞结果的各种变化有关。在这项研究中，从曼彻斯特收集的柴油废气颗粒物(DEPS)、城市粉尘标准参考物质SRM1649a和空气中三种不同的PM样本，在无细胞测试中测试了它们氧化DNA、增加细胞内ROS水平和诱导哺乳动物细胞中CYP1A1基因表达的能力。此外，分别采用3-(4，5-dimethylthiazol-2-yl)-2，5-diphenyltetrazolium溴化法和碱性彗星试验对PM的细胞毒性和遗传毒性进行了评价。所有PM样品都在无细胞实验中催化了Fenton反应，但只有DEP在哺乳动物细胞中通过二氯二氢荧光素二醋酸酯氧化测定导致ROS的产生。然而，没有证据表明ROS的增加是多环芳烃代谢的结果，因为城市粉尘、曼彻斯特粉尘样本而不是DEP诱导的CYP1A1表达。城市粉尘对小鼠胚胎成纤维细胞(MEF)的细胞毒性比其他PM样本更强，并且在没有S9激活的GreenScreen人类细胞检测中也诱导了GADD45a的表达，这表明存在直接作用的基因毒物。通过碱性彗星试验评估，城市粉尘和DEP在MEF中产生的DNA损伤水平与曼彻斯特PM所致水平更高总而言之，细胞毒性和遗传毒性检测的结果并不能证明ROS的产生是PM毒性的唯一决定因素。这表明有机成分对这种毒性有很大的贡献，需要进一步的工作来更好地表征ROS和有机成分对PM诱导的毒性的贡献程度。
Ambient air particulate matter (PM)-associated reactive oxygen species (ROS) have been linked to a variety of altered cellular outcomes. In this study, three different PM samples from diesel exhaust particles (DEPs), urban dust standard reference material SRM1649a and air collected in Manchester have been tested for their ability to oxidise DNA in a cell-free assay, to increase intracellular ROS levels and to induce CYP1A1 gene expression in mammalian cells. In addition, the cytotoxicity and genotoxicity of PM were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and alkaline comet assay, respectively. All PM samples catalysed the Fenton reaction in a cell-free assay, but only DEP resulted in the generation of ROS as measured by dichlorodihydrofluorescein diacetate oxidation in mammalian cells. However, there was no evidence that increased ROS was a consequence of polycyclic aromatic hydrocarbon metabolism via CYP1A1 induction as urban dust, the Manchester dust samples but not DEP-induced CYP1A1 expression. Urban dust was more cytotoxic in murine embryonic fibroblasts (MEFs) than the other PM samples and also induced expression of GADD45a in the GreenScreen Human Cell assay without S9 activation suggesting the presence of a direct-acting genotoxicant. Urban dust and DEP produced comparable levels of DNA damage, as assessed by the alkaline comet assay, in MEFs at higher levels than those induced by Manchester PM. In conclusion, results from the cytotoxic and genotoxic assays are not consistent with ROS production being the sole determinant of PM-induced toxicity. This suggests that the organic component can contribute significantly to this toxicity and that further work is required to better characterise the extent to which ROS and organic components contribute to PM-induced toxicity.