微小RNA-223-3p调控Notch通路对屋尘螨所致过敏性鼻炎小鼠鼻腔炎症反应的影响实验研究OACSTPCD
Effect of microRNA-223-3p on nasal inflammation in mice with allergic rhinitis caused by house dust mite via regulating Notch pathway
目的:探讨微小RNA(miR)-223-3p对屋尘螨(HDM)所致过敏性鼻炎(AR)小鼠鼻腔炎症反应的影响及相关机制.方法:80只BALB/c小鼠随机分为对照组、HDM致AR组(AR组)、miR-223-3p激动剂组(agomiR-223-3p组)、miR-223-3p 抑制剂组(antagomiR-223-3p 组)及 miR-223-3p 激动剂+敲低 Notch1 组(agomiR-223-3p+sh-Notch1组),每组各16只.利用HDM变应原提取物构建AR小鼠模型,并给予miR-223-3p激动剂(miR-223-3p agomir)、miR-223-3p 抑制剂(miR-223-3p antagomir)及敲低 Notch1 的腺相关病毒(AAV-sh-Notch1)滴鼻治疗.末次激发后,对小鼠进行AR症状评估,并收集血清、鼻腔灌洗液(NALF)和鼻黏膜组织标本.HE染色检测鼻黏膜损伤.Diff-Quik染色检测NALF中嗜酸性粒细胞数量.ELISA检测血清HDM特异性免疫球蛋白E(HDM-sIgE)和NALF中炎症因子水平.RT-qPCR检测鼻黏膜miR-223-3p、炎症因子及Notch通路相关mRNA水平.Western blot检测鼻黏膜Notch1、Jagged1蛋白水平.结果:与对照组比较,AR组小鼠鼻痒、喷嚏、流鼻涕情况严重,鼻黏膜增厚,可见大量炎症浸润;AR症状评分、血清HDM-sIgE水平、鼻黏膜miR-223-3p水平升高;NALF中嗜酸性粒细胞增多,白介素(IL)-4、IL-5、IL-13水平升高,IL-12、干扰素-γ(IFN-γ)水平降低;鼻黏膜IL-4、IL-5、IL-13 mRNA和Notch1、Jagged1 mRNA 及蛋白水平升高,IL-12、IFN-γ mRNA 水平降低(均P＜0.05).agomiR-223-3p 组上述炎症反应较AR组增强,并激活Notch通路;antagomiR-223-3p组上述炎症反应较AR组减轻,并阻断Notch通路(均P＜0.05).与agomiR-223-3p组比较,敲低Notch表达可逆转miR-223-3p对AR小鼠鼻腔炎症反应的激活作用.结论:miR-223-3p通过激活Notch通路增强HDM所致AR小鼠鼻腔炎症反应.
Objective:To explore the effect of microRNA(miR-223-3p)on nasal inflammation induced by house dust mite(HDM)in mice with allergic rhinitis(AR)and its related mechanism.Methods:Eighty BALB/c mice were randomly divided into control group,HDM-induced AR group(AR group),miR-223-3p agonist group(agomiR-223-3p group),miR-223-3p inhibitor group(antagomiR-223-3p group)and miR-223-3p agonist+Notch1 knockdown group(agomiR-223-3p+sh-Notch1 group),16 mice in each group.HDM allergen extract was used to construct AR mice model,and miR-223-3p agonist(miR-223-3p agomir),miR-223-3p inhibitor(miR-223-3p antagomir)and adeno-associated virus of knockdown Notch1(AAV-sh-Notch1)nasal administration were used for treatment.After the last stimulation,AR symptom of mice was performed.Serum,nasal lavage fluid(NALF)and nasal mucosal specimens were collected.HE staining was used to detect nasal mucosal damage.Diff Quik staining was used to detect the number of eosinophils in NALF.ELISA was used to detect HDM specific IgE(HDM-sIgE)level in serum and inflammatory factors levels in NALF.RT-qPCR was used to detect miR-223-3p,inflammatory fac-tors and Notch pathway related mRNA levels in nasal mucosa.Western blot was used to detect Notch1 and Jagged1 protein levels in nasal mucosa.Results:Compared with the control group,the nasal itchiness,sneezing and runny nose of mice in AR group were serious,and the nasal mucosa was thickened and a large number of inflammatory infiltrates could be seen.AR symptom score,serum HDM-sIgE level and nasal mucosa miR-223-3p level were increased.In NALF,eosinophils increased,interleukin(IL)-4,IL-5 and IL-13 levels increased,and IL-12 and interferon-y(IFN-y)levels decreased.The levels of IL-4,IL-5 and IL-13 mRNA,Notch1 and Jagged1 mRNA and protein in nasal mucosa were increased,while the levels of IL-12 and IFN-γ mRNA were decreased(all P＜0.05).Compared with AR group,agomiR-223-3p group enhanced the above inflammatory response and activated Notch pathway.In antagomiR-223-3p group,the above inflammatory response was alleviated and Notch pathway was blocked(all P＜0.05).Compared with agomiR-223-3p group,Notch down-expression can reverse the activation of miR-223-3p on nasal inflammatory response in AR mice.Conclusion:MiR-223-3p enhances nasal inflammatory response in HDM induced AR mice by ac-tivating the Notch pathway.
汪奕龙;宋杰;吴湘明
海南医学院第二附属医院耳鼻咽喉头颈外科,海南海口 570311
过敏性鼻炎微小RNA-223-3pNotch通路屋尘螨炎症反应小鼠
Allergic rhinitismicroRNA-223-3pNotch pathwayHouse dust miteInflammation responseMice
《陕西医学杂志》 2024 (005)
598-603,609 / 7
海南省卫生健康行业科研项目(22A200265)
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