Department of Otorhinolaryngology Head and Neck Surgery, the Second Affiliated Hospital of Nanchang University, Nanchang, P. R. China.
Am J Rhinol Allergy. 2022 Jul;36(4):510-520. doi: 10.1177/19458924221086059. Epub 2022 Apr 11.
Allergic rhinitis (AR) is an allergic disease of nasal mucosa. LncRNAs are key modulators affecting AR development. Neverthelss, the impact of lncRNA ANRIL in AR is not clear.
This work decided to study the mechanism underlying the impact of ANRIL on TLR4 expression through targeting miR-16-5p during autophagy and epithelial barrier dysfunction in the progression of AR.
Human nasal epithelial cells were exposed to TNF-α to establish AR cell model, AR mice model was constructed by ovalbumin (OVA) treatment. QRT-PCR or western blot assays were applied to measure the levels of mRNA and proteins. Dual-luciferase reporter gene detection and RIP assay were conducted to verify the association between ANRIL and miR-16-5p. Autophagy flux assessment by mRFP-GFP-LC3 method was performed to detect autophagy level.
AR progression could induce the autophagy, and the expressions of tight junction proteins were downregulated in AR cell model. Moreover, knockdown of ANRIL reversed the effect of AR on autophagy-related protein and tight junction proteins MiR-16-5p was found to be bound with ANRIL and miR-16-5p inhibitor could reverse ANRIL knockdown-induced downregulation of autophagy-related proteins and epithelial barrier dysfunction. In addition, miR-16-5p directly targeted TLR4. Furthermore, knockdown of ANRIL reversed miR-16-5p and TLR4 expression, autophagy level, and tight junction protein levels in nasal mucosa of AR mice.
This study illustrated that ANRIL acted as a promotion factor in AR induced autophagy and epithelial barrier dysfunction by enhancing the expression of TLR4 via interacting with miR-16-5p.
变应性鼻炎（AR）是一种鼻黏膜的过敏性疾病。lncRNAs 是影响 AR 发展的关键调节因子。然而，lncRNA ANRIL 在 AR 中的作用尚不清楚。
本研究旨在探讨 ANRIL 通过靶向自噬和上皮屏障功能障碍过程中的 miR-16-5p 影响 TLR4 表达的机制，从而在 AR 的进展中发挥作用。
采用 TNF-α 处理人鼻上皮细胞建立 AR 细胞模型，采用卵清蛋白（OVA）处理构建 AR 小鼠模型。应用 qRT-PCR 或 Western blot 检测 mRNA 和蛋白水平。采用双荧光素酶报告基因检测和 RIP 实验验证 ANRIL 与 miR-16-5p 的关系。采用 mRFP-GFP-LC3 方法检测自噬流来评估自噬水平。
AR 进展可诱导自噬，AR 细胞模型中紧密连接蛋白表达下调。此外，敲低 ANRIL 可逆转 AR 对自噬相关蛋白和紧密连接蛋白的作用。发现 miR-16-5p 与 ANRIL 结合，miR-16-5p 抑制剂可逆转 ANRIL 敲低诱导的自噬相关蛋白和上皮屏障功能障碍下调。此外，miR-16-5p 可直接靶向 TLR4。进一步研究发现，敲低 ANRIL 可逆转 AR 小鼠鼻黏膜中 miR-16-5p 和 TLR4 表达、自噬水平以及紧密连接蛋白水平。
本研究表明，ANRIL 通过增强与 miR-16-5p 的相互作用来上调 TLR4 的表达，从而在 AR 诱导的自噬和上皮屏障功能障碍中发挥促进作用。
