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Cytotoxicity and toxicoproteomic analyses of human lung epithelial cells exposed to extracts of atmospheric particulate matters on PTFE filters using acetone and water.
Ecotoxicology and Environmental Safety ( IF 6.8 ) Pub Date : 2020-01-25 , DOI: 10.1016/j.ecoenv.2020.110223 Zhi-Jie Tang 1 , Zhao-Ming Cao 1 , Xue-Wen Guo 1 , Hong-Juan Chen 2 , Yi Lian 3 , Wei-Juan Zheng 2 , Yi-Jun Chen 1 , Hong-Zhen Lian 1 , Xin Hu 1
Ecotoxicology and Environmental Safety ( IF 6.8 ) Pub Date : 2020-01-25 , DOI: 10.1016/j.ecoenv.2020.110223 Zhi-Jie Tang 1 , Zhao-Ming Cao 1 , Xue-Wen Guo 1 , Hong-Juan Chen 2 , Yi Lian 3 , Wei-Juan Zheng 2 , Yi-Jun Chen 1 , Hong-Zhen Lian 1 , Xin Hu 1
Affiliation
Differences of cytotoxicity associated with exposure to different extracts of atmospheric particulate matters (PMs) are still not well characterized by in vitro toxicoproteomics. In this study, in vitro cytotoxicity assays and toxicoproteomic analyses were carried out to investigate toxic effects of PM collected using polytetrafluoroethylene (PTFE) filters extracted with acetone for PM2.1 and water for PM2.1 and PM10 on A549 human lung epithelial cells. The cytotoxicity assays based on cell viability, cell apoptosis and reactive oxygen species generation indicated that PM2.1 extracted with acetone had the highest toxicity. iTRAQ labeling and LC-MS/MS analyses indicated that the number of differentially expressed proteins in A549 cells affected by PM2.1 extracted with acetone was noticeably higher than that of the other two groups. Hierarchical cluster analyses showed that the influences of the extracts of PM2.1 and PM10 using water on the proteome of A549 cells were similar, whereas significantly different from the effect of PM2.1 extracted with acetone. Pathways analyses indicated that PM2.1 extracted with acetone influenced the expression of proteins involved in 14 pathways including glycolysis/gluconeogenesis, pentose phosphate pathway, proteasome, etc. PM2.1 extracted with water affected the expression of proteins involved in 3 pathways including non-homologous end-joining, ribosome and endocytosis. However, PM10 extracted with water affected the expression of proteins involved in only spliceosome pathway. The extracts of PM using different extractants to detach PM from PTFE filters influenced the cytotoxic effects of PM and the proteome of A549 cells. Therefore, extractants should be assessed carefully before the investigations on cytotoxicity to improve the compatibility of experimental results among research teams.
中文翻译:
使用丙酮和水在PTFE过滤器上暴露于大气颗粒物提取物的人肺上皮细胞的细胞毒性和毒理学分析。
体外毒物组学仍不能很好地表征与暴露于大气颗粒物(PMs)的不同提取物相关的细胞毒性差异。在这项研究中,进行了体外细胞毒性测定和毒理学分析,以研究用丙酮萃取的聚四氟乙烯(PTFE)滤器收集的PM对PM2.1以及水对PM2.1和PM10收集的PM对A549人肺上皮细胞的毒性作用。基于细胞活力,细胞凋亡和活性氧生成的细胞毒性试验表明,丙酮提取的PM2.1具有最高的毒性。iTRAQ标记和LC-MS / MS分析表明,丙酮提取的PM2.1影响的A549细胞中差异表达蛋白的数量明显高于其他两组。层次聚类分析表明,用水提取PM2.1和PM10对A549细胞蛋白质组的影响相似,而与用丙酮提取PM2.1的影响却有显着差异。途径分析表明,丙酮提取的PM2.1影响糖酵解/糖异生,磷酸戊糖途径,蛋白酶体等14种途径的蛋白质表达。水提取的PM2.1影响3种途径的蛋白质表达,包括非同源末端连接,核糖体和胞吞作用。但是,用水提取的PM10仅影响剪接体途径中涉及的蛋白质表达。使用不同的萃取剂将PM从PTFE过滤器中分离出来的PM提取物影响PM和A549细胞蛋白质组的细胞毒性作用。因此,
更新日期:2020-01-26
中文翻译:
使用丙酮和水在PTFE过滤器上暴露于大气颗粒物提取物的人肺上皮细胞的细胞毒性和毒理学分析。
体外毒物组学仍不能很好地表征与暴露于大气颗粒物(PMs)的不同提取物相关的细胞毒性差异。在这项研究中,进行了体外细胞毒性测定和毒理学分析,以研究用丙酮萃取的聚四氟乙烯(PTFE)滤器收集的PM对PM2.1以及水对PM2.1和PM10收集的PM对A549人肺上皮细胞的毒性作用。基于细胞活力,细胞凋亡和活性氧生成的细胞毒性试验表明,丙酮提取的PM2.1具有最高的毒性。iTRAQ标记和LC-MS / MS分析表明,丙酮提取的PM2.1影响的A549细胞中差异表达蛋白的数量明显高于其他两组。层次聚类分析表明,用水提取PM2.1和PM10对A549细胞蛋白质组的影响相似,而与用丙酮提取PM2.1的影响却有显着差异。途径分析表明,丙酮提取的PM2.1影响糖酵解/糖异生,磷酸戊糖途径,蛋白酶体等14种途径的蛋白质表达。水提取的PM2.1影响3种途径的蛋白质表达,包括非同源末端连接,核糖体和胞吞作用。但是,用水提取的PM10仅影响剪接体途径中涉及的蛋白质表达。使用不同的萃取剂将PM从PTFE过滤器中分离出来的PM提取物影响PM和A549细胞蛋白质组的细胞毒性作用。因此,