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暴露于水醇萃煤尘提取物的肺细胞中的氧化应激和与炎症、DNA 损伤及金属暴露相关的基因表达改变。

Oxidative stress and alterations in the expression of genes related to inflammation, DNA damage, and metal exposure in lung cells exposed to a hydroethanolic coal dust extract.

机构信息

Environmental and Computational Chemistry Group, School of Pharmaceutical Sciences, University of Cartagena, Zaragocilla Campus, 130014, Cartagena, Colombia.

Functional Toxicology Group, School of Pharmaceutical Sciences, University of Cartagena, Zaragocilla Campus, 130014, Cartagena, Colombia.

出版信息

Mol Biol Rep. 2022 Jun;49(6):4861-4871. doi: 10.1007/s11033-022-07341-0. Epub 2022 Mar 25.

Abstract

BACKGROUND

Open cast mining is well known as a concerning source of environmental and public health problems. This work aimed to obtain a hydroethanolic coal dust extract (≤ 38 µm) and to characterize its composition with particular regard to content of organic compounds by GC/MS, as well as describe its toxicity in vitro on Calu-1 after exposure to several concentrations (0-500 μg/mL).

MATERIALS AND RESULTS

Cytotoxicity was measured with MTT assay and DCFH-DA probe was employed to estimate the amount of reactive oxygen species (ROS) in Calu-1 cells. RT-PCR was employed to quantify relative expression of genes associated with inflammation, oxidative stress, as well as metals, and lipid metabolism. Seventeen organic compounds were identified in the extract, highlighting undecane, dodecane, pentadecane and benzo[a]anthracene, 6,12-dimethyl-1,2,3,4-tetrahydro-. Cytotoxicity test showed a decrease trend in the cell viability after 24 h hours from the concentration of 62.5 µg/mL. Further, the extract raised intracellular ROS when compared with control. Expression levels of CYP1A1, IL-8, IL-6, MT1X, and NQO1 were up-regulated when cells were exposed to 125 µg/mL of coal dust, whereas PPAR-α was down-regulated, likely involving aryl hydrocarbon receptor regulation.

CONCLUSIONS

In short, this study shows that despite hydroethanolic coal dust extract is not cytotoxic to Calu-1 cells, it produces an elevation of intracellular ROS and alters the expression in marker genes of oxidative stress, inflammation, metal transport, xenobiotic and lipid metabolism. These findings suggest that chemicals present in coal dust are biologically active and may interfere key biochemical process in the living organisms.

摘要

背景

露天开采是众所周知的环境和公共卫生问题的来源。本工作旨在获得水醇煤粉提取物(≤38μm),并通过 GC/MS 特别关注其有机化合物含量来对其进行成分分析,以及描述其在暴露于不同浓度(0-500μg/mL)的情况下对 Calu-1 的体外毒性。

材料和结果

通过 MTT 测定法测量细胞毒性,并使用 DCFH-DA 探针估计 Calu-1 细胞中活性氧(ROS)的含量。采用 RT-PCR 定量与炎症、氧化应激以及金属和脂质代谢相关的基因的相对表达。在提取物中鉴定出 17 种有机化合物,突出显示十一烷、十二烷、十五烷和苯并[a]蒽、6,12-二甲基-1,2,3,4-四氢-。细胞毒性试验显示,在 62.5μg/mL 浓度下,细胞活力在 24 小时后呈下降趋势。此外,与对照相比,提取物增加了细胞内 ROS。当细胞暴露于 125μg/mL 的煤粉时,CYP1A1、IL-8、IL-6、MT1X 和 NQO1 的表达水平上调,而 PPAR-α 下调,可能涉及芳烃受体调节。

结论

总之,本研究表明,尽管水醇煤粉提取物对 Calu-1 细胞没有细胞毒性,但它会导致细胞内 ROS 升高,并改变氧化应激、炎症、金属转运、外来物和脂质代谢标志物基因的表达。这些发现表明煤粉中的化学物质具有生物活性,可能会干扰生物体内的关键生化过程。

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