Chang Sandra, Linderholm Angela, Franzi Lisa, Kenyon Nicholas, Grasberger Helmut, Harper Richart
Department of Internal Medicine, Division of Pulmonary and Critical Care Medicine, School of Medicine, University of California at Davis, Davis, CA 95616, USA.
Department of Medicine, University of Michigan, Ann Arbor, MI 48109, USA.
Free Radic Biol Med. 2013 Dec;65:38-46. doi: 10.1016/j.freeradbiomed.2013.06.012. Epub 2013 Jun 11.
Enhanced reactive oxygen species production in allergic airways is well described and correlates with increased airway contractions, inflammatory cell infiltration, goblet cell metaplasia, and mucus hypersecretion. There is also an abundance of interleukin-4/interleukin-13 (IL-4/IL-13)- or interleukin-5-secreting cells that are thought to be central to the pathogenesis of allergic asthma. We postulated that the dual oxidases (DUOX1 and DUOX2), members of the nicotinamide adenine dinucleotide phosphate oxidase family that release hydrogen peroxide (H2O2) in the respiratory tract, are critical proteins in the pathogenesis of allergic airways. DUOX activity is regulated by cytokines, including IL-4 and IL-13, and DUOX-mediated H2O2 influences several important features of allergic asthma: mucin production, IL-8 secretion, and wound healing. The objective of this study was to establish the contribution of DUOXs to the development of allergic asthma in a murine model. To accomplish this goal, we utilized a DUOXA-deficient mouse model (Duoxa(-/-)) that lacked maturation factors for both DUOX1 and DUOX2. Our results are the first to demonstrate evidence of DUOX protein and DUOX functional activity in murine airway epithelium. We also demonstrate that DUOXA maturation factors are required for airway-specific H2O2 production and localization of DUOX to cilia of fully differentiated airway epithelial cells. We compared wild-type and Duoxa(-/-) mice in an ovalbumin exposure model to determine the role of DUOX in allergic asthma. In comparison to DUOX-intact mice, Duoxa(-/-) mice had reduced mucous cell metaplasia and lower levels of TH2 cytokine levels in bronchoalveolar fluid. In addition, increased airway resistance in response to methacholine was observed in Duoxa(+/+) mice, as expected, but was absent in Duoxa(-/-) mice. Surprisingly, Duoxa(-/-) mice had decreased influx of neutrophils in bronchoalveolar fluid and lung tissue sections associated with a lower level of the chemotactic cytokine IL-6. These findings suggest that DUOX-derived H2O2 has an important role in signaling neutrophils into allergic airways.
过敏性气道中活性氧生成增强已有充分描述,且与气道收缩增加、炎性细胞浸润、杯状细胞化生和黏液分泌亢进相关。此外,还存在大量分泌白细胞介素-4/白细胞介素-13(IL-4/IL-13)或白细胞介素-5的细胞,这些细胞被认为是过敏性哮喘发病机制的核心。我们推测,双氧化酶(DUOX1和DUOX2)是烟酰胺腺嘌呤二核苷酸磷酸氧化酶家族成员,可在呼吸道释放过氧化氢(H2O2),是过敏性气道发病机制中的关键蛋白。DUOX活性受细胞因子调节,包括IL-4和IL-13,且DUOX介导的H2O2影响过敏性哮喘的几个重要特征:黏蛋白产生、IL-8分泌和伤口愈合。本研究的目的是在小鼠模型中确定DUOX对过敏性哮喘发展的作用。为实现这一目标,我们利用了DUOXA缺陷小鼠模型(Duoxa(-/-)),该模型缺乏DUOX1和DUOX2的成熟因子。我们的结果首次证明了小鼠气道上皮中DUOX蛋白和DUOX功能活性的证据。我们还证明,气道特异性H2O2生成以及DUOX定位于完全分化的气道上皮细胞的纤毛需要DUOXA成熟因子。我们在卵清蛋白暴露模型中比较了野生型和Duoxa(-/-)小鼠,以确定DUOX在过敏性哮喘中的作用。与DUOX完整的小鼠相比,Duoxa(-/-)小鼠的黏液细胞化生减少,支气管肺泡灌洗液中TH2细胞因子水平降低。此外,正如预期的那样,Duoxa(+/+)小鼠对乙酰甲胆碱的气道阻力增加,但Duoxa(-/-)小鼠没有。令人惊讶的是,Duoxa(-/-)小鼠支气管肺泡灌洗液和肺组织切片中中性粒细胞的流入减少,趋化细胞因子IL-6水平较低。这些发现表明,DUOX衍生的H2O2在将中性粒细胞信号传导至过敏性气道中起重要作用。