House dust mite-induced asthma causes oxidative damage and DNA double-strand breaks in the lungs
House dust mite-induced asthma causes oxidative damage and DNA double-strand breaks in the lungs
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DOI：
10.1016/j.jaci.2016.02.017
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发表时间：
2016-07-01
影响因子：
14.2
通讯作者：
Engelward, Bevin P.
中科院分区：
文献类型：
--
作者：
Chan, Tze Khee;Loh, Xin Yi;Engelward, Bevin P.
背景：哮喘与气道炎症和氧化应激有关。高水平的活性氧和氮物质可诱导细胞毒性DNA损伤。然而，很少有人知道过敏原诱导的DNA损伤和DNA修复作为哮喘相关pathology.Objective调制器的可能作用：我们试图研究DNA损伤和DNA损伤反应诱导的屋尘螨（HDM）在体内和体外。方法：我们测量DNA双链断裂（DSB），DNA修复蛋白，和细胞凋亡在anHDM诱导的过敏性哮喘模型和哮喘患者的肺标本。为了研究DNA修复，我们用DSB修复抑制剂NU 7441处理小鼠。为了研究HDM对人支气管上皮细胞的直接DNA损伤作用，我们将BEAS-2B细胞暴露于HDM，并测量DNA损伤和活性氧水平。结果：HDM挑战增加了肺蛋白质（3-硝基酪氨酸），脂质（8-异前列腺素）和核酸（8-氧代鸟嘌呤）的氧化损伤水平。HDM诱导的DNA DSB的免疫组织化学证据通过支气管上皮中DSB标志物γ组蛋白2AX（H2 AX）病灶水平的增加来揭示。暴露于HDM的BEAS-2B细胞显示出增强的DNA损伤，如通过使用彗星试验和γ H2 AX染色所测量的。在人类哮喘患者的肺组织中，我们观察到DNA修复蛋白和细胞凋亡水平的增加，如caspase-3切割，caspase激活的DNase水平，和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记染色所示。值得注意的是，NU 7441增强了DNA损伤和细胞因子的产生在支气管上皮细胞和细胞凋亡的过敏性气道，牵连DSBs作为一个潜在的驱动程序的哮喘pathophysiology.Conclusion：这项工作呼吁注意活性氧和氮物种和HDM诱导的细胞毒性和DNA修复作为哮喘相关的病理生理学的调制器的潜在作用。
Background: Asthma is related to airway inflammation and oxidative stress. High levels of reactive oxygen and nitrogen species can induce cytotoxic DNA damage. Nevertheless, little is known about the possible role of allergen-induced DNA damage and DNA repair as modulators of asthma-associated pathology.Objective: We sought to study DNA damage and DNA damage responses induced by house dust mite (HDM) in vivo and in vitro.Methods: We measured DNA double-strand breaks (DSBs), DNA repair proteins, and apoptosis in anHDM-induced allergic asthma model and in lung samples from asthmatic patients. To study DNA repair, we treated mice with the DSB repair inhibitor NU7441. To study the direct DNA-damaging effect of HDM on human bronchial epithelial cells, we exposed BEAS-2B cells to HDM and measured DNA damage and reactive oxygen species levels.Results: HDM challenge increased lung levels of oxidative damage to proteins (3-nitrotyrosine), lipids (8-isoprostane), and nucleic acid (8-oxoguanine). Immunohistochemical evidence for HDM-induced DNA DSBs was revealed by increased levels of the DSB marker gamma Histone 2AX (H2AX) foci in bronchial epithelium. BEAS-2B cells exposed to HDM showed enhanced DNA damage, as measured by using the comet assay and gamma H2AX staining. In lung tissue from human patients with asthma, we observed increased levels of DNA repair proteins and apoptosis, as shown by caspase-3 cleavage, caspase-activated DNase levels, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling staining. Notably, NU7441 augmented DNA damage and cytokine production in the bronchial epithelium and apoptosis in the allergic airway, implicating DSBs as an underlying driver of asthma pathophysiology.Conclusion: This work calls attention to reactive oxygen and nitrogen species and HDM-induced cytotoxicity and to a potential role for DNA repair as a modulator of asthma-associated pathophysiology.