15-脂氧合酶 1 在鼻息肉中通过细胞外信号调节激酶的激活促进 CCL26/eotaxin 3 的表达。
15-Lipoxygenase 1 in nasal polyps promotes CCL26/eotaxin 3 expression through extracellular signal-regulated kinase activation.
机构信息
Department of Otolaryngology-Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai; University of Pittsburgh Asthma Institute@UPMC, Department of Environmental and Occupational Health, Graduate School of Public Health, Pittsburgh, Pa.
Department of Otolaryngology-Head and Neck Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; University of Pittsburgh Asthma Institute@UPMC, Department of Environmental and Occupational Health, Graduate School of Public Health, Pittsburgh, Pa.
出版信息
J Allergy Clin Immunol. 2019 Nov;144(5):1228-1241.e9. doi: 10.1016/j.jaci.2019.06.037. Epub 2019 Jul 11.
BACKGROUND
15-Lipoxygenase 1 (15LO1) is expressed in airway epithelial cells in patients with type 2-high asthma in association with eosinophilia. Chronic rhinosinusitis with nasal polyps (CRSwNP) is also associated with type 2 inflammation and eosinophilia. CCL26/eotaxin 3 has been reported to be regulated by 15LO1 in lower airway epithelial cells. However, its relation to 15LO1 in patients with CRSwNP or mechanisms for its activation are unclear.
OBJECTIVE
We sought to evaluate 15LO1 and CCL26 expression in nasal epithelial cells (NECs) from patients with CRSwNP and healthy control subjects (HCs) and determine whether 15LO1 regulates CCL26 in NECs through extracellular signal-regulated kinase (ERK) activation.
METHODS
15LO1, CCL26, and phosphorylated ERK were evaluated in NECs from patients with CRSwNP and HCs. 15LO1/CCL26 and CCL26/cytokeratin 5 were colocalized by means of immunofluorescence. IL-13-stimulated NECs were cultured at an air-liquid interface with or without 15-lipoxygenase 1 gene (ALOX15) Dicer-substrate short interfering RNAs (DsiRNA) transfection, a specific 15LO1 enzymatic inhibitor, and 2 ERK inhibitors. Expression of 15LO1 and CCL26 mRNA and protein was analyzed by using quantitative RT-PCR, Western blotting, and ELISA.
RESULTS
15LO1 expression was increased in nasal polyp (NP) epithelial cells compared with middle turbinate epithelial cells from patients with CRSwNP and HCs. 15LO1 expression correlated with CCL26 expression and colocalized with CCL26 expression in basal cells of the middle turbinate and NPs from patients with CRSwNP. In primary NECs in vitro, IL-13 induced 15LO1 and CCL26 expression. 15LO1 knockdown and inhibition decreased IL-13-induced ERK phosphorylation and CCL26 expression. ERK inhibition (alone) similarly decreased IL-13-induced CCL26. Phosphorylated ERK expression was increased in NECs from CRSwNP subjects and positively correlated with both 15LO1 and CCL26 expression.
CONCLUSIONS
15LO1 expression is increased in NP epithelial cells and contributes to CCL26 expression through ERK activation. 15LO1 could be considered a novel therapeutic target for CRSwNP.
背景
15-脂氧合酶 1（15LO1）在 2 型高哮喘患者的气道上皮细胞中表达，与嗜酸性粒细胞增多有关。慢性鼻-鼻窦炎伴鼻息肉（CRSwNP）也与 2 型炎症和嗜酸性粒细胞增多有关。CCL26/嗜酸性粒细胞趋化因子 3 已被报道在低气道上皮细胞中受 15LO1 调节。然而，其与 CRSwNP 患者中 15LO1 的关系或其激活的机制尚不清楚。
目的
我们旨在评估 CRSwNP 患者和健康对照（HC）的鼻上皮细胞（NEC）中 15LO1 和 CCL26 的表达，并确定 15LO1 是否通过细胞外信号调节激酶（ERK）激活来调节 NEC 中的 CCL26。
方法
评估 CRSwNP 患者和 HC 的 NEC 中的 15LO1、CCL26 和磷酸化 ERK。通过免疫荧光共定位 15LO1/CCL26 和 CCL26/细胞角蛋白 5。在空气-液界面培养经 IL-13 刺激的 NEC，并进行 15-脂氧合酶 1 基因（ALOX15）Dicer 底物短发夹 RNA（DsiRNA）转染、特异性 15LO1 酶抑制剂和 2 种 ERK 抑制剂的处理。通过定量 RT-PCR、Western 印迹和 ELISA 分析 15LO1 和 CCL26 mRNA 和蛋白的表达。
结果
与 CRSwNP 患者和 HC 的中鼻甲上皮细胞相比，鼻息肉（NP）上皮细胞中 15LO1 的表达增加。15LO1 的表达与 CCL26 的表达相关，并与 CRSwNP 患者中鼻甲和 NP 中的 CCL26 表达的基底细胞共定位。在体外原代 NEC 中，IL-13 诱导 15LO1 和 CCL26 的表达。15LO1 敲低和抑制减少了 IL-13 诱导的 ERK 磷酸化和 CCL26 的表达。ERK 抑制（单独）也同样降低了 IL-13 诱导的 CCL26。CRSwNP 患者的 NEC 中磷酸化 ERK 的表达增加，并与 15LO1 和 CCL26 的表达呈正相关。
结论
NP 上皮细胞中 15LO1 的表达增加，并通过 ERK 激活促进 CCL26 的表达。15LO1 可被视为 CRSwNP 的一种新的治疗靶点。
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