Autophagy regulates DUOX1 localization and superoxide production in airway epithelial cells during chronic IL-13 stimulation.
Autophagy regulates DUOX1 localization and superoxide production in airway epithelial cells during chronic IL-13 stimulation.
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自噬调节慢性IL-13刺激时呼吸道上皮细胞DUOX1的定位和超氧化物的产生。
DOI：
10.1016/j.redox.2017.09.013
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发表时间：
2018-04
期刊：
影响因子：
11.4
通讯作者：
Brody SL
中科院分区：
文献类型：
--
作者：
Dickinson JD;Sweeter JM;Warren KJ;Ahmad IM;De Deken X;Zimmerman MC;Brody SL
气道上皮是与环境的广泛界面，要求精心策划的反应以适当地调节炎症。经典地，自噬是响应于外部细胞应激而触发的稳态途径，并且在慢性气道疾病中升高。最近的研究结果强调了自噬在囊泡运输和蛋白质分泌中的额外作用，暗示了自噬途径在疾病中复杂的细胞反应中的作用。Th 2细胞因子IL-13和IL-4在哮喘和其他气道疾病中增加，导致慢性炎症。以前，我们观察到IL-13以自噬依赖的方式增加气道上皮细胞中的活性氧（ROS）。在这里，我们测试了我们的假设，即自噬是IL-13介导的通过NADPH氧化酶DUOX 1产生超氧化物所必需的。使用由OVA过敏原诱导的Th 2介导的炎症的小鼠模型，我们观察到IL-13和IL-4的肺量升高，伴随着增加的自噬体水平，通过LC 3BII蛋白水平和免疫染色确定。在OVA攻击的肺中，ROS水平升高，DUOX 1表达增加70倍。为了解决自噬和ROS在气道上皮中的作用，我们用IL-13或IL-4处理原代人气管支气管上皮细胞。延长7天的治疗增加了自噬体的形成和降解，而短暂的激活没有影响。在平行培养条件下，IL-13和IL-4增加细胞内超氧化物水平，如通过电子顺磁共振（EPR）光谱测定的。延长的IL-13激活增加DUOX 1，定位于顶膜。通过siRNA沉默DUOX 1减弱了IL-13介导的超氧化物的增加，但没有降低自噬活性。值得注意的是，自噬调节蛋白ATG 5的消耗显著降低了超氧化物，而没有降低总DUOX 1水平。然而，耗尽的ATG 5，减少DUOX 1定位在顶膜。研究结果表明，非典型的自噬活性调节DUOX 1依赖的定位所需的细胞内超氧化物的产生在Th 2炎症。因此，在慢性Th 2炎症性气道疾病中，自噬蛋白可能是持续细胞内超氧化物产生的原因。气道上皮细胞是Th 2炎症过程中ROS的重要来源。IL-13/IL-4细胞因子增加气道上皮中的超氧化物和自噬。IL-13介导的细胞内超氧化物的增加是DUOX 1依赖性的。IL-13介导的超氧化物水平的增加依赖于自噬。自噬是正确的细胞内DUOX 1定位所必需的。
The airway epithelium is a broad interface with the environment, mandating well-orchestrated responses to properly modulate inflammation. Classically, autophagy is a homeostatic pathway triggered in response to external cellular stresses, and is elevated in chronic airway diseases. Recent findings highlight the additional role of autophagy in vesicle trafficking and protein secretion, implicating autophagy pathways in complex cellular responses in disease. Th2 cytokines, IL-13 and IL-4, are increased in asthma and other airway diseases contributing to chronic inflammation. Previously, we observed that IL-13 increases reactive oxygen species (ROS) in airway epithelial cells in an autophagy-dependent fashion. Here, we tested our hypothesis that autophagy is required for IL-13-mediated superoxide production via the NADPH oxidase DUOX1. Using a mouse model of Th2-mediated inflammation induced by OVA-allergen, we observed elevated lung amounts of IL-13 and IL-4 accompanied by increased autophagosome levels, determined by LC3BII protein levels and immunostaining. ROS levels were elevated and DUOX1 expression was increased 70-fold in OVA-challenged lungs. To address the role of autophagy and ROS in the airway epithelium, we treated primary human tracheobronchial epithelial cells with IL-13 or IL-4. Prolonged, 7-day treatment increased autophagosome formation and degradation, while brief activation had no effect. Under parallel culture conditions, IL-13 and IL-4 increased intracellular superoxide levels as determined by electron paramagnetic resonance (EPR) spectroscopy. Prolonged IL-13 activation increased DUOX1, localized at the apical membrane. Silencing DUOX1 by siRNA attenuated IL-13-mediated increases in superoxide, but did not reduce autophagy activities. Notably, depletion of autophagy regulatory protein ATG5 significantly reduced superoxide without diminishing total DUOX1 levels. Depletion of ATG5, however, diminished DUOX1 localization at the apical membrane. The findings suggest non-canonical autophagy activity regulates DUOX1-dependent localization required for intracellular superoxide production during Th2 inflammation. Thus, in chronic Th2 inflammatory airway disease, autophagy proteins may be responsible for persistent intracellular superoxide production. The airway epithelium is a significant source of ROS during Th2 inflammation. IL-13/IL-4 cytokines increase both superoxide and autophagy in airway epithelium. IL-13-mediated increase in intracellular superoxide is DUOX1 dependent. The IL-13-mediated increase in superoxide levels depends on autophagy. Autophagy is required for proper intracellular DUOX1 localization.
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