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水中挥发性有机化合物混合物的允许最大污染水平对黑腹果蝇总DNA、抗氧化基因表达及核糖体DNA序列的影响。

Effects of permissible maximum-contamination levels of VOC mixture in water on total DNA, antioxidant gene expression, and sequences of ribosomal DNA of Drosophila melanogaster.

作者信息

Doganlar Oguzhan, Doganlar Zeynep Banu, Tabakcioglu Kiymet

机构信息

Faculty of Medicine, Department of Medical Biology, Trakya University, Edirne, Turkey.

出版信息

Environ Sci Pollut Res Int. 2015 Oct;22(20):15610-20. doi: 10.1007/s11356-015-4741-y. Epub 2015 May 28.

Abstract

In this study, we aimed to investigate the mutagenic and carcinogenic potential of a volatile organic compound (VOC) mixture with references to the response of D.melanogaster using selected antioxidant gene expressions, RAPD assay and base-pair change of ribosomal 18S, and the internal transcribed spacer, ITS2 rDNA gene sequences. For this purpose, Drosophila melanogaster Oregon R, reared under controlled conditions on artificial diets, were treated with the mixture of thirteen VOCs, which are commonly found in water in concentrations of 10, 20, 50, and 75 ppb for 1 and 5 days. In the random amplified polymorphic DNA (RAPD) assay, band changes were clearly detected, especially at the 50 and 75 ppb exposure levels, for both treatment periods, and the band profiles exhibited clear differences between the treated and untreated flies with changes in band intensity and the loss/appearance of bands. Quantitative real-time PCR (qRT-PCR) analysis of Mn-superoxide dismutase (Mn-SOD), catalase (CAT) and glutathione-synthetase (GS) expressions demonstrated that these markers responded significantly to VOC-induced oxidative stress. Whilst CAT gene expressions increased linearly with increasing concentrations of VOCs and treatment times, the 50- and 75-ppb treatments caused decreases in GS expressions compared to the control at 5 days. Treatment with VOCs at both exposure times, especially in high doses, caused gene mutation of the 18S and the ITS2 ribosomal DNA. According to this research, we thought that the permissible maximum-contamination level of VOCs can cause genotoxic effect especially when mixed.

摘要

在本研究中,我们旨在通过选定的抗氧化基因表达、随机扩增多态性DNA(RAPD)分析以及核糖体18S和内部转录间隔区ITS2核糖体DNA基因序列的碱基对变化,研究挥发性有机化合物(VOC)混合物的致突变和致癌潜力,并参考黑腹果蝇的反应。为此,在人工饲料上于受控条件下饲养的黑腹果蝇俄勒冈R品系,用13种VOC的混合物进行处理,这些VOC在水中的常见浓度为10、20、50和75 ppb,处理1天和5天。在随机扩增多态性DNA(RAPD)分析中,在两个处理时间段内,尤其是在50和75 ppb的暴露水平下,均能清楚地检测到条带变化,并且条带图谱显示,处理组和未处理组果蝇之间在条带强度变化以及条带的缺失/出现方面存在明显差异。对锰超氧化物歧化酶(Mn-SOD)、过氧化氢酶(CAT)和谷胱甘肽合成酶(GS)表达进行的定量实时PCR(qRT-PCR)分析表明,这些标志物对VOC诱导的氧化应激有显著反应。虽然CAT基因表达随VOC浓度和处理时间的增加呈线性增加,但在5天时,50和75 ppb处理组的GS表达与对照组相比有所下降。在两个暴露时间用VOC处理,尤其是高剂量处理,会导致18S和ITS2核糖体DNA发生基因突变。根据这项研究,我们认为VOC的允许最大污染水平可能会导致遗传毒性效应,尤其是在混合时。

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