局部变应性鼻炎小鼠模型的建立与研究

A murine model of local allergic rhinitis

目的 建立卵清蛋白（ovalbumin,OVA）诱导的小鼠局部变应性鼻炎（local allergic rhinitis,LAR）和变应性鼻炎（AR）模型,并探讨LAR与AR的区别与联系.方法将30只BALB/c小鼠分为5组,每组6只,分别为Al组（鼻部致敏10 d组）、A1对照组、A2组（鼻部致敏25 d组）、A2对照组、B组（腹腔致敏组）.A组以小剂量OVA（对照组以磷酸盐缓冲液PBS替代OVA）鼻部致敏和激发建立模型;B组常规腹腔注射OVA和氢氧化铝致敏,经鼻部OVA激发建立经典的AR模型.统计分析各组小鼠滴鼻激发后喷嚏数,以及苏木精-伊红染色后观察小鼠气道改变和鼻中隔嗜酸粒细胞计数;酶联免疫吸附试验测定鼻腔灌洗液中白细胞介素（IL）4、5、13和血清中OVA特异性IgE抗体（OVA-sIgE）浓度.使用Graph Pad Prism 6统计软件对数据进行分析处理.结果 鼻部致敏组A1组第10天小鼠血清sIgE处于其对照组水平（t=0.697,P＞0.05）,但出现明显的喷嚏症状和鼻中隔嗜酸粒细胞浸润,表现为人类LAR症状,鼻部致敏组A2组第25天血清中slgE为（2.710＆#177;1.406） ng/ml,相比其对照组的（0.221 ＆#177;0.080） ng/ml明显升高（=4.329,P＜0.05）,并伴气道重塑;鼻部致敏组A1组第10天鼻腔灌洗液中IL-5、IL-13较第25天（A2组）增高明显（t值分别为2.442、2.804,P值均＜0.05）.结论 短期OVA滴鼻建立的LAR小鼠表现为鼻局部产生大量IL-5、IL-13,但血清sIgE正常.持续的抗原接触诱发LAR小鼠血清sIgE升高,并伴发气道改变.

Objective To establish the murine models of local allergic rhinitis （LAR） and allergic rhinitis （AR） by using ovalbumin （OVA）,and to investigate the relationship between them.Methods Thirty BALB/c mice were divided into 5 groups,（1） the nasally sensitized group （group A1） that was challenged with OVA by a 10 d procedure,（2） the control group of A1 that was challenged with phosphatebuffered saline （PBS）,（3） the nasally sensitized group （group A2） that was challenged with OVA by a 25 d procedure,（4） the control group of A2 that was challenged with PBS,（5） the intraperitoneally sensitized group （group B）.The numbers of sneezing after final challenge were counted,and the serum OVA-specific immunoglobulin E （OVA-slgE）,interleukin （IL）-4,IL-13,IL-5 levels in nasal lavage fluid were measured by ELISA.Hematoxylin-eosin staining was performed to evaluate the histological change of nose and lung tissues.Graph Pad Prism 6 software was used to analyze the data.Results Nasally sensitized group A1 displayed LAR symptoms of sneezing and eosinophilic infiltrating,but without increased OVA-sIgE in serum on day 10 compared with the control group of A1（t =0.697,P 〉 0.05）,OVA-sIgE in serum of group A2（2.710 ＆#177; 1.406）ng/ml reached to statistical significance and with airway remodeling on day 25 compared with the control group of A2 （（0.221 ＆#177; 0.080） ng/ml,t =4.329,P 〈 0.05）.IL-5 and IL-13 in nasal fluid showed a significant increase in the nasally sensitized group A1,compared with the group A2 （t values were 2.442,2.804,P values were less then 0.05）.Conclusions A short time intranasal instillation with OVA could establish LAR murine model,continuing OVA challenge could increase serum sIgE level and with airway remodeling.LAR mice show a unique characteristic by expressing higher IL-5 and IL-13 in nose than AR mice,but sIgE in serum remains at a normal level.