sEH通过内质网应激通路调节COPD气道炎症的研究

Airway inflammation plays central role in the pathogenesis of chronic obstructive pulmonary disease (COPD).Cigarette smoke can cause airway inflammation involved in endoplasmic reticulum stress. In our previous study, we found that smoke stimulation up-regulate soluble epoxide hydrolase (sEH) then decrease the concentrations of epoxyeicosatrienoic acids which suppresses cigarette smoke extract-induced apoptosis in lung epithelial cells by inhibiting endoplasmic reticulum stress..In this study, we will exam the role of sEH on airway inflammation in sEH gene knockout mice by cigarette smoke induced COPD. In vitro, we will use sEH-shRNA and AAV-sEH or a selective epoxygenase inhibitor to test the role of sEH on inflammation in airway epithelial cells and the chemotaxis of macrophages. Moreover, we further test endoplasmic reticulum-resident kinase (PERK), eukaryotic initiation factor 2 (eIF2α), inositol-requiringenzyme 1α (IRE1α), and c-jun NH2-terminal kinase JNK pathway will be tested, and blocking endoplasmic reticulum stress pathway to found whether sEH involved airway inflammation by regulating the endoplasmic reticulum stress. We also exam the levels of EET / EHET, sEH metabolites, and the changes of endoplasmic reticulum Ca2+-ATPase (SERCA) to explore the molecular signaling mechanism involing endoplasmic reticulum stress induced inflammation.Our study will provide a new experimental basis for the pathogenesis of COPD, and provide possible new drugs for clinical treatment of COPD.

气道炎症是慢性阻塞性肺病（COPD）发病核心机制。烟雾刺激可引起气道细胞内质网应激参与气道炎症。我们前期发现，烟雾刺激可上调可溶性表氧化物水解酶(sEH)减少花生四烯酸经表氧化物代谢产物(EET)，减少内质网应激诱导的上皮细胞凋亡参与COPD。.本研究拟利用烟熏诱导小鼠COPD及sEH基因敲除模型，观察sEH对气道炎症细胞及炎症因子的变化；体外培养气道上皮细胞，通过sEH干扰、转染技术及气道-巨噬细胞共培养技术，研究sEH对气道细胞炎症因子及巨噬细胞趋化作用；观察肺组织及气道上皮细胞内质网结构及内质网应激主要信号PERK/eIF2α及IRE1/JNK的变化，检测EET及sEH代谢产物DHET含量及内质网Ca2+-ATP酶，并阻断内质网信号观察对气道细胞炎症影响，探讨sEH参与气道炎症可能机制。本研究为阐明sEH在COPD气道炎症中的作用提供理论依据，为COPD防治提供新药物靶点。

慢性阻塞性肺疾病（COPD）的核心发病机制是慢性气道炎症，本研究通过在动物模型和细胞水平研究了sEH对香烟烟雾暴露下COPD气道炎症调节作用及分子机制研究。研究发现，sEH促进了COPD发病，sEH基因敲除对COPD发病有抑制作用，表现在能够改善气道炎症反应、气道粘液高分泌和肺气肿。进一步发现，sEH敲除能够改善气道上皮细胞内质网结构及抑制内质网应激信号GRP78、IRE1、PERK的表达，sEH对内质网Ca2+-ATP酶的影响仅在细胞水平能减少其蛋白的表达，提示sEH可能部分是通过内质网Ca2+-ATP酶发挥作用；sEH敲除还能够促进肺部巨噬细胞由M1型向M2型转化，减轻炎症因子的释放；sEH敲除参与气道炎症反应是通过抑制IRE1 /JNK/AP-1信号通路发挥作用。本研究首次发现sEH在介导COPD气道炎症的作用，为进一步研发新药奠定了基础。