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首页> 外文会议>Annual conference of the International Society of Exposure Science >Differences in biological activity among household dust samples detected by changes in expression of biomarker genes in zebrafish Danio rerio
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Differences in biological activity among household dust samples detected by changes in expression of biomarker genes in zebrafish Danio rerio

机译：斑马鱼Danio rerio中生物标志物基因表达的变化检测到的家庭灰尘样品中生物活性的差异

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Household dust is a complex mixture of particulates with sorbed chemicals. Since assessment of all complex mixture substances by analytical chemistry is not practical, this study investigated relative biological activity among dust samples by in vivo assessment of biomarkers of toxicant exposure with the zebrafish Danio rerio model. Samples were collected from vacuum cleaners of 6 households in Edinburgh UK, sieved to < 63 μm and extracted using n-hexane:acetone (3:1). Three independent replicates of zebrafish larvae were exposed to each dust extract, spiked positive controls, or vehicle controls. Total RNA extracted from the larvae was reverse transcribed to generate cDNA for quantitative real-time PCR. Relative changes in gene expression of cyp 1a (a biomarker of oxidative stress and metabolism of toxicants such as polycyclic aromatic hydrocarbons) and vtg (a biomarker of exposure to estrogenic substances) were compared among dust samples and positive controls. Significant induction of cypla (p<0.05) was observed in two household dust extracts compared to the other extracts and the vehicle control. Although the spiked positive control induced vtg expression, there was no induction of vtg transcripts with exposure to un-spiked samples. Results indicate the presence of organic toxicants in some households at sufficient quantities to induce cypla activity, and that households differ in the presence of bioactive substances associated with dust. While numerous substances present within households are suspected of having estrogenic activity, none of the dust samples had sufficient activity to induce vtg expression in larval zebrafish. This study demonstrates that the zebrafish model is an effective screening tool to identify biological activity in household dust samples. Dust samples with biological activity can be prioritized for further examination of compounds present, or for examination of factors that contribute to biological activity within specific households.
机译：家庭灰尘是微粒与吸附的化学物质的复杂混合物。由于通过分析化学方法评估所有复杂的混合物质是不切实际的，因此本研究通过使用斑马鱼Danio rerio模型体内评估有毒物质暴露的生物标志物来研究粉尘样品中的相对生物活性。从英国爱丁堡的6户家庭的真空吸尘器中收集样品，筛分至<63μm，然后使用正己烷：丙酮（3：1）进行提取。斑马鱼幼虫的三个独立复制品分别暴露于每种粉尘提取物，加标的阳性对照或媒介物对照。从幼虫提取的总RNA被逆转录生成cDNA，用于实时定量PCR。比较了粉尘样品和阳性对照中cyp 1a（氧化应激的生物标志物和多环芳香烃等有毒物质代谢的生物标志物）和vtg（暴露于雌激素物质的生物标志物）基因表达的相对变化。与其他提取物和赋形剂对照相比，在两种家用粉尘提取物中观察到了Cycy的显着诱导（p <0.05）。尽管加标的阳性对照诱导了vtg表达，但暴露于未加标的样品并没有诱导vtg转录物。结果表明，某些家庭中有足够数量的有机毒物引起胞浆活动，并且家庭中与灰尘相关的生物活性物质也存在差异。尽管怀疑家庭中存在的许多物质具有雌激素活性，但没有粉尘样品具有足够的活性来诱导幼虫斑马鱼中的vtg表达。这项研究表明，斑马鱼模型是一种有效的筛选工具，可以识别家庭灰尘样品中的生物活性。可以对具有生物活性的粉尘样品进行优先排序，以进一步检查存在的化合物，或检查特定家庭中有助于生物活性的因素。

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