Institut Universitaire de Cardiologie et de Pneumologie de Québec; and.
Groupe de Recherche en Ecologie Buccale, Faculté de médecine dentaire, Université Laval, Québec City, Québec, Canada.
Am J Respir Cell Mol Biol. 2024 Nov;71(5):603-616. doi: 10.1165/rcmb.2024-0089OC.
MicroRNA (miR)-155-5p increases in innate and adaptive immune cells in response to IL-13 and is associated with the severity of asthma. However, little is known about its role in airway structural cells. Bronchial epithelial cells (BECs) isolated from healthy donors and patients with severe asthma were stimulated with IL-13. miR-155-5p expression and release were measured by real-time (RT)-PCR in BECs and in their derived exosomes. Modulation of miR-155-5p in BECs was performed using transfection of miR-155-5p inhibitor and mimic. IL-13 receptor α1 (IL-13Rα1), IL-13Rα2, MUC5AC, IL-8, and eotaxin-1 expression was measured by RT-PCR and Western blot analysis. The BEC repair process was assessed by a wound-healing assay. IL-13Rα1 and IL-13Rα2 expression and downstream pathways were evaluated by Western blot analysis. A dual luciferase assay was used to identify miR-155-5p target genes associated with IL-13R signaling. BECs from patients with severe asthma showed increased expression and exosomal release of miR-155-5p at baseline with amplification by IL-13 stimulation. BECs from patients with asthma expressed more IL-13Rα1 and less IL-13Rα2 than those from healthy donors, and IL-13Rα1 but not IL-13Rα2 induced miR-155-5p expression under IL-13 stimulation. miR-155-5p overexpression favored , , and through the IL-13Rα1/SOCS1/STAT6 pathway while delaying the repair process by downregulating IL-13Rα2/MAPK14/c-Jun/c-fos signaling. The dual luciferase assay confirmed that miR-155-5p modulates both IL-13R pathways by directly targeting SOCS1, c-fos, and MAPK14. miR-155-5p is overexpressed in BECs from patients with severe asthma and regulates IL-13Rα1 and IL-13Rα2 expression and signaling, favoring expression of mucin- and eosinophil-related genes to the detriment of airway repair. These results show that miR-155-5p may contribute to airway epithelial cell dysfunction in patients with severe asthma.
微小 RNA（miR）-155-5p 在先天和适应性免疫细胞中响应白细胞介素-13（IL-13）而增加，并与哮喘的严重程度相关。然而，关于其在气道结构细胞中的作用知之甚少。从健康供体和严重哮喘患者中分离出的支气管上皮细胞（BEC），并使用白细胞介素-13（IL-13）刺激。通过实时（RT）-PCR 测量 BEC 及其衍生的外泌体中的 miR-155-5p 表达和释放。通过转染 miR-155-5p 抑制剂和模拟物来调节 BEC 中的 miR-155-5p。使用 RT-PCR 和 Western blot 分析测量 IL-13 受体 α1（IL-13Rα1）、IL-13Rα2、MUC5AC、IL-8 和嗜酸性粒细胞趋化因子-1（eotaxin-1）的表达。通过划痕愈合试验评估 BEC 修复过程。通过 Western blot 分析评估 IL-13Rα1 和 IL-13Rα2 表达和下游途径。使用双荧光素酶测定法鉴定与 IL-13R 信号相关的 miR-155-5p 靶基因。严重哮喘患者的 BEC 在基线时表现出 miR-155-5p 的表达和外泌体释放增加，并且在 IL-13 刺激下扩增。与健康供体相比，哮喘患者的 BEC 表达更多的 IL-13Rα1 和更少的 IL-13Rα2，并且只有 IL-13Rα1 而不是 IL-13Rα2 在 IL-13 刺激下诱导 miR-155-5p 表达。miR-155-5p 过表达通过 IL-13Rα1/SOCS1/STAT6 通路促进 、 和 ，同时通过下调 IL-13Rα2/MAPK14/c-Jun/c-fos 信号转导来延迟修复过程。双荧光素酶测定法证实 miR-155-5p 通过直接靶向 SOCS1、c-fos 和 MAPK14 来调节两种 IL-13R 途径。miR-155-5p 在严重哮喘患者的 BEC 中过度表达，并调节 IL-13Rα1 和 IL-13Rα2 的表达和信号转导，有利于粘蛋白和嗜酸性粒细胞相关基因的表达，从而损害气道修复。这些结果表明，miR-155-5p 可能导致严重哮喘患者的气道上皮细胞功能障碍。
