陈旭青 马华安 周龙云 严道南 刘书芬 吴继勇

中圖分类号 R765.2;R285.5 文献标志码 A 文章编号 1001-0408（2021）10-1187-09

DOI 10.6039/j.issn.1001-0408.2021.10.06

摘 要 目的：初步研究黄芪甲苷对变应性鼻炎（AR）模型小鼠改善作用的可能机制。方法：将C57/BL6小鼠随机分为空白组、模型组和黄芪甲苷组，每组10只。除空白组外，其余大鼠均于第0、7、14、21～27天以卵清蛋白为致敏源进行攻击以复制AR模型。于造模第15～27天，黄芪甲苷组小鼠按0.02 mL/g腹腔注射黄芪甲苷40 mg/kg（第21～27天均于攻击致敏前1 h给药），空白组和模型组小鼠腹腔注射等体积生理盐水，每天1次。末次攻击致敏24 h后，观察各组小鼠鼻黏膜组织中炎症细胞的浸润情况，检测其鼻腔灌洗液中白细胞介素4（IL-4）、IL-5、干扰素γ（IFN-γ）含量，鼻黏膜和脾脏组织中活性氧簇（ROS）水平和磷酸化Janus激酶2（p-JAK2）、磷酸化信号转导及转录激活蛋白6（p-STAT6）阳性细胞计数，以及脾脏组织中JAK2、STAT6蛋白的磷酸化水平（即p-JAK2/JAK2比值、p-STAT6/STAT6比值）。结果：与空白组比较，模型组小鼠鼻黏膜组织中炎症细胞（嗜酸粒细胞和肥大细胞）计数，鼻腔灌洗液中IL-4、IL-5含量，鼻黏膜和脾脏组织中ROS水平和p-JAK2、p-STAT6阳性细胞计数，脾脏组织中p-JAK2/JAK2比值、p-STAT6/STAT6比值均显著升高（P<0.05），鼻腔灌洗液中INF-γ含量显著降低（P<0.05）。与模型组比较，黄芪甲苷组小鼠鼻黏膜组织中炎症细胞计数，鼻腔灌洗液中IL-4、IL-5含量，鼻黏膜和脾脏组织中ROS水平和p-JAK2、p-STAT6阳性细胞数量，脾脏组织中p-JAK2/JAK2比值、p-STAT6/STAT6比值均显著降低（P<0.05），鼻腔灌洗液中INF-γ含量显著增加（P<0.05）。结论：黄芪甲苷能够有效改善AR模型小鼠变应性炎症反应，其机制可能与下调JAK2/STAT6信号通路及ROS水平有关。

关键词 变应性鼻炎;黄芪甲苷;活性氧簇;Janus激酶2/信号转导及转录激活蛋白6信号通路;小鼠

Preliminary Study on the Improvement Effects of Astragaloside Ⅳ on Allergic Rhinitis Model Mice

CHEN Xuqing1，MA Huaan1，ZHOU Longyun2，YAN Daonan1，LIU Shufen3，WU Jiyong1（1. Dept. of Otorhinolaryngology， the Affiliated Hospital of Nanjing University of TCM， Nanjing 210029， China; 2. Dept. of Rehabilitation， the First Affiliated Hospital of Nanjing Medical University， Nanjing 210029， China; 3. Faculty of Basic Medicine， Longhua Hospital Affiliated to Shanghai University of TCM， Shanghai 210032， China）

ABSTRACT   OBJECTIVE： To preliminarily study the potential mechanism of astragaloside Ⅳ on allergic rhinitis （AR） model mice. METHODS： C57/BL6 mice were randomly divided into blank group， model group and astragaloside Ⅳ group， with 10 mice in each group. Except for blank group， AR model was prepared by sensitization and challenge with ovalbumin on day 0， 7， 14 and 21-27.  Astragaloside Ⅳ group was given astragaloside Ⅳ 40 mg/kg intraperitoneally at the dose of 0.02 mL/g on the 15th to 27th day of modeling （given the drug 1 h before challenge sensitization on the 21st to 27th day）. Blank group and model group were given constant volume of normal saline intraperitoneally， once a day. Twenty-four hours after sensitization from the last challenge， the infiltration of inflammatory cells in the nasal mucosa of each group was observed， and the contents of interleukin 4 （IL-4）， IL-5 and interferon gamma （IFN-γ） in the nasal lavage fluid were measured. The levels of reactive oxygen species （ROS）， and the count of phosphorylated Janus kinase 2 （p-JAK2） and phosphorylation signal transduction and activation of transcription protein 6 （p-STAT6） positive cells in the nasal mucosa and spleen as well as the phosphorylation levels of JAK2 and STAT6 proteins in spleen tissue （i.e. p-JAK2/JAK2 ratio， p-STAT6/STAT6 ratio） were also determined. RESULTS： Compared with blank group， the number of inflammatory cells in the nasal mucosa （eosinophils and mast cells） in the model group， the contents of IL-4 and IL-5 in the nasal lavage fluid， and the levels of ROS in the nasal mucosa and spleen tissues in the model group， the count of p-JAK2 and p-STAT6 positive cells increased significantly， the p-JAK2/JAK2 ratio， p-STAT6/STAT6 ratio in the spleen tissue were significantly increased （P<0.05）， and the content of INF-γ in the nasal lavage fluid was significantly decreased （P<0.05）. Compared with model group， the count of inflammatory cells infiltrated in the nasal mucosa， the contents of IL-4 and IL-5 in the nasal cavity lavage fluid， the level of ROS and the number of p-JAK2 and p-STAT6 positive cellsin the nasal mucosa and spleen tissue as well as the p-JAK2/JAK2 ratio and p-STAT6/STAT6 ratio in spleen tissue were decreased significantly （P<0.05）， and the content of INF-γ in nasal lavage fluid was significantly increased （P<0.05）.  CONCLUSIONS： Astragaloside Ⅳ can effectively improve the inflammatory response in AR model mice， the mechanism of which may be related to down-regulation of JAK2/STAT6 signaling pathway and ROS level.

KEYWORDS   Allergic rhinitis; Astragaloside Ⅳ; ROS;JAK2/STAT6 signaling pathway; Mice

变应性鼻炎（AR）为耳鼻咽喉头颈外科最常见的疾病之一[1-3]。据统计，世界范围内约有10.0%～40.4%的人口具有AR的临床表现[4-6]。研究表明，内源性活性氧簇（ROS）水平升高以及Janus激酶2（JAK2）、信号转导及转录激活蛋白6（STAT6）信号分子活化是变应性炎症发生发展的重要病理机制，而选择性降低ROS水平，抑制JAK2、STAT6信号分子活化，则能显著改善AR等变应性炎症[7-10]。……