Protocols to Measure Oxidative Stress and DNA Damage in Asthma.
Protocols to Measure Oxidative Stress and DNA Damage in Asthma.
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测量哮喘氧化应激和 DNA 损伤的方案。
DOI：
10.1007/978-1-0716-2364-0_22
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Sur,Sanjiv
中科院分区：
文献类型：
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作者：
Hosoki,Koa;Chakraborty,Anirban;Hazra,TapasK;Sur,Sanjiv
哮喘与包括DNA在内的生物分子的氧化应激和氧化损伤有关。在这里，我们描述了在小鼠过敏性气道炎症模型中量化活性氧（ROS）和氧化应激标志物的方案。我们还提供了详细的方法来测量DNA损伤的长期实时PCR DNA损伤定量（LORD-Q）测定和基因特异性DNA损伤分析的长扩增子(LA)-qPCR。此外，我们描述了用质谱法定量肺基因组DNA中氧化DNA碱基病变的方法，以及测量8-氧鸟嘌呤DNA糖基酶（OGG1）的酶活性的方法。利用这些方法，可以阐明过敏性炎症和哮喘中的氧化应激和DNA损伤水平。
Asthma is associated with oxidative stress and oxidative damage of biomolecules, including DNA. Here, we describe the protocols to quantify reactive oxygen species (ROS) and oxidative stress markers in a mouse model of allergic airway inflammation. We also provide detailed methods to measure DNA damage by long-run real-time PCR for DNA-damage quantification (LORD-Q) assay and gene-specific DNA damage analyses by long amplicon (LA)-qPCR. Additionally, we describe methods to quantify oxidized DNA base lesions in lung genomic DNA by mass spectrometry, and to measure enzymatic activity of 8-oxoguanine DNA glycosylase (OGG1). Using these methods, the levels of oxidative stress and DNA damage in allergic inflammation and asthma can be elucidated.