14,15-Epoxyeicosatrienoic acid suppresses cigarette smoke condensate-induced inflammation in lung epithelial cells by inhibiting autophagy
14,15-Epoxyeicosatrienoic acid suppresses cigarette smoke condensate-induced inflammation in lung epithelial cells by inhibiting autophagy
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DOI：
10.1152/ajplung.00161.2016
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发表时间：
2016-11-01
影响因子：
4.9
通讯作者：
Wang, Haoyan
中科院分区：
文献类型：
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作者：
Li, Yunxiao;Yu, Ganggang;Wang, Haoyan
环氧二十碳三烯酸（Epoxyeicosatrienoic Acids，ESTs）是游离花生四烯酸的代谢产物，由细胞色素P-450（CYP 450）环氧合酶产生。雌二醇具有抗炎、抗凋亡和抗氧化活性。然而，依地平对香烟烟雾诱导的肺部炎症的影响尚不清楚。自噬被认为参与了慢性阻塞性肺疾病的发病机制。此外，核红细胞相关因子2（Nrf 2）是一种调节许多抗氧化基因的转录因子，被认为调节多种肺部疾病的抗氧化防御。此外，已经报道了Ehrs、自噬和Nrf 2之间的相互作用。本研究的目的是探讨14，15-EET对香烟烟雾冷凝物（CSC）诱导的人支气管上皮细胞系（Beas-2B）炎症的影响，并确定其潜在机制是否涉及通过抑制自噬来调节Nrf 2。通过Western印迹分析评估自噬和自噬信号通路蛋白（LC 3B、p62、PI 3 K、Akt、p-Akt和p-mTOR）和抗炎蛋白（Nrf 2和HO-1）的表达。通过腺病毒mRFP-GFP-LC 3转染检测自噬体和自溶酶体。ELISA法检测炎症因子（IL-6、IL-8和MCP-1）。使用携带p62短发夹RNA的慢病毒载体来干扰p62表达以评估p62对Nrf 2表达的影响。免疫细胞化学法检测Nrf 2的表达。14，15-EET处理导致IL-6、IL-8和MCP-1分泌显著减少，Nrf 2积累和HO-1表达增加。此外，14，15-EET抑制CSC诱导的Beas-2B细胞自噬。14，15-EET的抗炎作用机制涉及抑制自噬和增加p62水平，随后Nrf 2易位到细胞核中，然后上调抗氧化酶HO-1的表达。14，15-EET通过抑制自噬促进Nrf 2的积累来保护CSC诱导的肺部炎症。
Epoxyeicosatrienoic acids (EETs) are metabolic products of free arachidonic acid, which are produced through cytochrome P-450 (CYP) epoxygenases. EETs have anti-inflammatory, antiapoptotic, and antioxidative activities. However, the effect of EETs on cigarette smoke-induced lung inflammation is not clear. Autophagy is believed to be involved in the pathogenesis of chronic obstructive pulmonary disease. In addition, nuclear erythroid-related factor 2 (Nrf2), a transcription factor that regulates many antioxidant genes, is thought to regulate antioxidant defenses in several lung diseases. In addition, interaction between EETs, autophagy, and Nrf2 has been reported. The aim of this study was to explore the effect of 14,15-EET on cigarette smoke condensate (CSC)-induced inflammation in a human bronchial epithelial cell line (Beas-2B), and to determine whether the underlying mechanisms involved in the regulation of Nrf2 through inhibition of autophagy. Autophagy and expression of autophagy signaling pathway proteins (LC3B, p62, PI3K, Akt, p-Akt, and p-mTOR) and anti-inflammatory proteins (Nrf2 and HO-1) were assessed via Western blot analysis. Autophagosomes and autolysosomes were detected by adenoviral mRFP-GFP-LC3 transfection. Inflammatory factors (IL-6, IL-8, and MCP-1) were detected by ELISA. Lentiviral vectors carrying p62 short hairpin RNA were used to interfere with p62 expression to evaluate the effect of p62 on Nrf2 expression. Nrf2 expression was determined through immunocytochemistry. 14,15-EET treatment resulted in a significant reduction in IL-6, IL-8, and MCP-1 secretion, and increased accumulation of Nrf2 and expression of HO-1. In addition, 14,15-EET inhibited CSC-induced autophagy in Beas-2B cells. The mechanism of the anti-inflammatory effect of 14,15-EET involved inhibition of autophagy and an increase in p62 levels, followed by translocation of Nrf2 into the nucleus, which then upregulated expression of the antioxidant enzyme HO-1. 14,15-EET protects against CSC-induced lung inflammation by promoting accumulation of Nrf2 via inhibition of autophagy.