miR-205-5p靶向HMGB1/TLR4途径对变应性鼻炎免疫功能的作用研究

Effect of miR-205-5p targeting HMGB1/TLR4 pathway on immune function in allergic rhinitis

摘要:

目的 研究微小RNA-205-5p （miR-205-5p）靶向高迁移率族蛋白B1/Toll样受体4（HMGB1/TLR4）途径改善变应性鼻炎（AR）免疫功能的作用。方法 选取60只雄性大鼠，分为正常对照组（A组）、变应性鼻炎组（B组）、变应性鼻炎+miR-205-5p agomir组（C组）、变应性鼻炎+miR-205-5p antagomir组（D组）共4组，每组各15只，A组不做任何处理，B、C、D组均采用卵清白蛋白鼻腔强化致敏建立AR模型。模型建立完成后，C组尾部注射300 μg miR-205-5p agomir，D组尾部注射miR-205-5p antagomir，A、B组均注射生理盐水。1周后评价各组行为学、免疫相关指标[干扰素-γ（IFN-γ）、白介素-2（IL-2）、白介素-4（IL-4）、白介素-10（IL-10）、免疫球蛋白E （IgE）]、鼻黏膜炎性反应、上皮细胞间紧密连接、内质网形态及紧密连接蛋白、HMGB1/TLR4途径蛋白表达量。miR-205-5p与TLR4的靶向关系使用双荧光素酶报告实验分析。结果 4组行为学评分、IL-4、IgE、HMGB1、TLR4、NF-κB表达量、紧密连接宽度从低到高分别为A组P均<0.05）。双荧光素酶报告结果表明miR-205-5p可与TLR4靶向结合。结论 miR-205-5p可靶向HMGB1/TLR4途径抑制Th1/Th2失衡所引起的AR免疫障碍，加强鼻黏膜屏障功能，从而减少变应原的入侵，缓解AR相关症状，对AR的治疗产生积极作用。

Abstract:

Objective To study the effect of microrna-205-5p (mir-205-5p) targeting high mobility group protein B1/toll like receptor 4 (HMGB1/TLR4) pathway on immune function of allergic rhinitis (AR).Methods Sixty male rats were selected and divided into normal control group (group A), AR group (group B), AR+miR-205-5p agomir group (group C), AR+miR-205-5p antagomir group (group D) with 15 rats in each group. The rats of group A did not receive any treatment, while those of group B, C, and D were sensitized with ovalbumin nasal enhancement sensitization to establish AR model. After model establishment, 300 μg of miR-205-5p agomir and miR-205-5p antagomir were injected into the tail of rats of group C and group D respectively, and normal saline into both group A and group B. One week later, the behavioral, immune-related indexes[including interferon-γ (IFN-γ), interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10), immunoglobulin E (IgE)], nasal mucosal inflammatory response, tight junctions between epithelial cells, endoplasmic reticulum morphology and expressions of tight junction proteins, HMGB1/TLR4 pathway proteins were evaluated. The targeting relationship between miR-205-5p and TLR4 was analyzed using a dual-luciferase reporter assay.Results The orders from lowest to highest of behavioral score, expression levels of IL-4, IgE, HMGB1, TLR4, and NF-κB as well as tight junction width were group A, group B, group C and group D. The orders from lowest to highest of expression levels of IFN-γ, IL-2, IL-10 and tight junction protein were group D, group C, group B and group A, and their differences among different groups were all statistically significant (all P<0.05).The dual-luciferase reporter results indicated that miR-205-5p could target TLR4.Conclusion miR-205-5p can target the HMGB1/TLR4 pathway to inhibit the AR immune disorder caused by Th1/Th2 imbalance, strengthen the barrier function of the nasal mucosa, thereby reducing the invasion of allergens, relieving AR-related symptoms, and having a positive effect on the treatment of AR.