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炭黑颗粒、柴油机尾气颗粒及城市空气颗粒物及其提取物对人肺泡上皮细胞系(A549)和人单核细胞系(THP-1)的遗传毒性作用。

Genotoxic effects of carbon black particles, diesel exhaust particles, and urban air particulates and their extracts on a human alveolar epithelial cell line (A549) and a human monocytic cell line (THP-1).

作者信息

Don Porto Carero A, Hoet P H, Verschaeve L, Schoeters G, Nemery B

机构信息

Environmental Toxicology, Vlaamse Instelling voor Technologisch Onderzoek (VITO), Mol, Belgium.

出版信息

Environ Mol Mutagen. 2001;37(2):155-63. doi: 10.1002/em.1023.

Abstract

The possible genotoxicity of small particulate matter has been under investigation for the last 10 years. Diesel exhaust particles (DEP) are considered as "probably carcinogenic" (IARC group 2A) and a number of studies show genotoxic effects of urban particulate matter (UPM). Carbon black (CB) is carcinogenic in rats. In this study the cytotoxic and genotoxic potency of these three particle types was investigated by exposing human cells (A549 and THP-1 cell lines) in vitro to CB, DEP (SRM 1650, NIST), and UPM (SRM 1648, NIST) for 48 hr. Cytotoxicity was assessed using the Alamar Blue assay, whereas genotoxicity was assessed using the single-cell gel electrophoresis (comet assay). The particles were characterized with regard to their mean diameter in tissue culture medium (CB 100 nm, DEP 400 nm, UPM 2 microm), their total carbon content (CB 99%, DEP 85%, UPM 15%), and their acid-soluble metal composition (UPM >> CB approximately DEP). The concentrations ranged from 16 ng/ml to 16 microg/ml for cytotoxicity tests and from 16 ng/ml to 1.6 microg/ml for genotoxicity tests. In both assays, paraquat was used as a reference chemical. The CB, DEP, and UPM particles showed no significant cytotoxicity. However, all three particles were able to cause significant DNA damage, although to a different extent in the two cell lines. The genotoxicity of washed particles and dichloromethane extracts was also investigated. In THP-1 cells CB washed particles and DEP extracts caused significant DNA damage. This difference in effect may be related to differences in size, structure, and composition of the particles. These results suggest that CB, DEP, and UPM are able to cause DNA damage and, therefore, may contribute to the causation of lung cancer. More detailed studies on influence of size, structure, and composition of the particles are needed.

摘要

在过去十年中,人们一直在研究小颗粒物可能具有的遗传毒性。柴油尾气颗粒(DEP)被视为“可能致癌”(国际癌症研究机构2A组),多项研究表明城市颗粒物(UPM)具有遗传毒性。炭黑(CB)对大鼠具有致癌性。在本研究中,通过在体外将人细胞(A549和THP-1细胞系)暴露于CB、DEP(美国国家标准与技术研究院标准参考物质1650)和UPM(美国国家标准与技术研究院标准参考物质1648)48小时,研究了这三种颗粒类型的细胞毒性和遗传毒性。使用阿拉玛蓝检测法评估细胞毒性,而使用单细胞凝胶电泳(彗星试验)评估遗传毒性。对颗粒的特征进行了如下测定:它们在组织培养基中的平均直径(CB为100纳米,DEP为400纳米,UPM为2微米)、总碳含量(CB为99%,DEP为85%,UPM为15%)以及酸溶性金属成分(UPM >> CB ≈ DEP)。细胞毒性试验的浓度范围为16纳克/毫升至16微克/毫升,遗传毒性试验的浓度范围为16纳克/毫升至1.6微克/毫升。在这两种检测中,百草枯用作参考化学品。CB、DEP和UPM颗粒均未显示出显著的细胞毒性。然而,尽管在两种细胞系中的程度不同,但所有三种颗粒都能够导致显著的DNA损伤。还研究了洗涤颗粒和二氯甲烷提取物的遗传毒性。在THP-1细胞中,CB洗涤颗粒和DEP提取物导致了显著的DNA损伤。这种效应差异可能与颗粒的大小、结构和组成差异有关。这些结果表明,CB、DEP和UPM能够导致DNA损伤,因此可能促成肺癌的发生。需要对颗粒的大小、结构和组成的影响进行更详细的研究。

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