小青龙汤对变应性鼻炎小鼠IL-33/ST2信号通路的影响  

Effect of Xiao Qinglongtang on IL-33/ST2 Signaling Pathway in Mice with Allergic Rhinitis

作　　者：张嘉骏 贺雪成 刘沁东 徐玮祯 闫亚杰 周敏 阮岩[2] ZHANG Jiajun;HE Xuecheng;LIU Qindong;XU Weizhen;YAN Yajie;ZHOU Min;RUAN Yan(First College of Clinical Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]广州中医药大学第一临床医学院,广州510006 [2]广州中医药大学第一附属医院,广州510004

出　　处：《中国实验方剂学杂志》2022年第14期13-19,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(8197151813);广东省基础与应用基础研究基金项目(2021A1515110444)。

摘　　要：目的:探讨小青龙汤对鸡卵清白蛋白(OVA)诱导的变应性鼻炎(AR)小鼠的治疗作用及对白细胞介素-33(IL-33)/IL-1受体样2(ST2)信号通路的影响。方法:72只雌性SPF级BALB/c小鼠随机分为正常组、模型组、氯雷他定组(2.05 mg·kg-1)、小青龙汤高、中、低剂量(20.02、10.01、5.005 g·kg-1)组。除正常组外,其余各组小鼠腹腔注射OVA溶液造成基础致敏,而后OVA溶液滴鼻激发致敏造成AR模型。每日局部滴鼻前30 min给药1次,正常组和模型组按20 mL·kg-1给与磷酸盐缓冲液(PBS),共7 d。记录末次OVA溶液滴鼻后10 min内小鼠打喷嚏和挠鼻的次数。给药7 d后收集血液,取小鼠鼻骨并脱钙制作病理切片。酶联免疫吸附测定法(ELISA)检测血清OVA特异性免疫球蛋白E(OVA-sIgE)、白细胞介素-4(IL-3)、白细胞介素-5(IL-4)、白细胞介素-13(IL-13)含量;苏木素-伊红(HE)染色观察鼻黏膜组织形态,糖原(PAS)染色观察鼻黏膜杯状细胞增生情况,吉姆萨(Giemsa)染色观察鼻黏膜嗜酸性粒细胞浸润情况;蛋白免疫印迹法(Western blot)检测鼻黏膜IL-33、ST2、IL-1受体辅助蛋白(IL-1RAP)蛋白表达水平。结果:与正常组比较,模型组小鼠打喷嚏,挠鼻次数显著升高(P<0.01),鼻黏膜水肿增厚,出现杯状细胞增生和嗜酸性粒细胞浸润,血清中OVA-sIgE、IL-4、IL-5、IL-13水平显著升高(P<0.01),鼻黏膜组织IL-33、ST2、IL-1RAP蛋白表达均有明显升高(P<0.05,P<0.01)。给药后,与模型组比较,高剂量小青龙汤组小鼠打喷嚏,挠鼻次数降低(P<0.01),鼻黏膜病理情况改善,血清中OVA-sIgE、IL-4、IL-5、IL-13水平降低(P<0.01),鼻黏膜组织IL-33、ST2、IL-1RAP蛋白的表达均有明显下降(P<0.05,P<0.01)。结论:小青龙汤对于OVA致敏AR小鼠有治疗作用,其作用可能与其调节IL-33/ST2信号通路及2型T辅助细胞(Th2)炎症因子,从而减轻Th2型免疫反应,缓解鼻黏膜损伤有关。Objective: To investigate the therapeutic effect of Xiao Qinglongtang(XQLT) on ovalbumin(OVA)-induced allergic rhinitis(AR) in mice and its effect on the interleukin-33(IL-33)/suppression of tumorigenicity 2(ST2)signaling pathway. Method: Seventy-two female BALB/c mice of SPF grade were randomly divided into a control group,a model group,a positive control group(loratadine,2.05 mg·kg-1),and low-,medium-,and high-dose(5.005,10.01,20.02 g·kg-1)XQLT groups. All mice except for those in the control group were sensitized by intraperitoneal injection of OVA solution,and the AR model was induced by intranasal drops of OVA solution. Thirty minutes before local intranasal drops,drugs were administered once,and mice in the control group and the model group received phosphate buffered saline(PBS)at 20 mL·kg;for 7 days. After the last intranasal drop of OVA solution,the times of sneezing and nasal rubbing of mice within 10 min was recorded. After drug administration for 7 days,blood samples were collected,and nasal bones of mice were decalcified for the preparation of pathological sections. The content of OVA-specific immunoglobulin E(OVA-sIgE),interleukin-4(IL-4),interleukin-5(IL-5),and interleukin-13(IL-13)was detected by enzyme-linked immunosorbent assay(ELISA)kits. Hematoxylin-eosin(HE)staining,periodic acid-Schiff(PAS)staining,and Giemsa staining were used to observe the pathological changes,goblet cell hyperplasia,and eosinophil infiltration of nasal mucosa,respectively. Western blot was used to detect the expression levels of IL-33,ST2,and IL-1 receptor accessory protein(IL-1RAP)in nasal mucosa. Result:Compared with the control group,the model group showed increased times of sneezing and nasal rubbing(P<0.01),edema and thickening of nasal mucosa,goblet cell hyperplasia and eosinophil infiltration,increased serum levels of OVA-sIgE,IL-4,IL-5 and IL-13(P<0.01),and increased protein expression of IL-33,ST2,and IL-1RAP in nasal mucosa(P<0.05,P<0.01). After drug administration,compared with the model group,the high-dose

关 键 词：变应性鼻炎 小青龙汤 2型T辅助细胞(Th2) 白细胞介素-33(IL-33) IL-1受体样2(ST2) 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33