小鼠嗜酸粒细胞性慢性鼻-鼻窦炎模型的建立  

Establishment of an eosinophilic chronic rhinosinusitis murine model

作　　者：卫红齐[1] 陈伟[2] 徐夏媛 刘志勇[1] 莫江伟 邢洪瑜 曹忠胜[1] WEI Hong-qi;CHEN Wei;XU Xia-yuan;LIU Zhi-yong;MO Jiang-wei;XING Hong-yu;CAO Zhong-sheng(Department of Otorhinolaryngology,the Second Affiliated Hospital of Soochow University,Suzhou 215004,Jian-gsu,China;Department of Otorhinolaryngology,General Hospital of Eastern Theater Command,PLA,Nanjing 210002,Jiangsu,China;Department of Otorhinolaryngology,Changshu Traditional Chinese Medicine Hospital,Changshu 215006,Jiangsu,China)

机构地区：[1]苏州大学附属第二医院耳鼻咽喉科,苏州215004 [2]东部战区总医院(原南京军区南京总医院)耳鼻咽喉科,南京210002 [3]苏州常熟市中医院新区医院耳鼻咽喉科,常熟215006

出　　处：《医学研究生学报》2022年第3期245-249,共5页Journal of Medical Postgraduates

基　　金：江苏省自然科学基金(BK20161388);苏州市科技发展计划(SKJY2021084)。

摘　　要：目的 嗜酸粒细胞性慢性鼻-鼻窦炎(ECRS)发病机制尚不明确。文中旨在探讨用普通抗原卵清蛋白(OVA)联合超抗原金黄色葡萄球菌肠毒素B(SEB)来诱导小鼠ECRS,建立小鼠ECRS实验模型。方法 将24只雌性小鼠随机分成4组:PBS组[腹腔注射及鼻腔滴入磷酸缓冲盐(PBS)]、SEB组(腹腔注射PBS,鼻腔滴入SEB)、OVA组(腹腔注射及鼻腔滴入OVA)及SEB+OVA组(腹腔注射OVA,鼻腔滴入SEB+OVA),每组6只。在d103获取小鼠鼻腔灌洗液(NLF),采用ELISA检测NLF中IFN-γ、IL-4、IL-5和IL-17A表达水平;HE染色观察小鼠鼻腔鼻窦黏膜息肉样病变及嗜酸性粒细胞(EOS)浸润情况;免疫组化检测嗜酸性粒细胞阳离子蛋白(ECP)表达水平。结果 OVA组和SEB+OVA组IL-4、IL-5浓度显著高于PBS组及SEB组(P<0.05);SEB+OVA组与OVA组比较,IL-5表达水平显著升高(P<0.05)。SEB+OVA组小鼠鼻腔鼻窦见息肉样病变伴明显EOS浸润,息肉样病变以及EOS计数均明显高于其他三组(P<0.05)。OVA组和SEB+OVA组鼻-鼻窦黏膜组织中均有较强表达ECP的阳性信号,表达ECP阳性细胞数明显多于PBS组及SEB组,且SEB+OVA组表达ECP阳性细胞数显著多于OVA组(P<0.05)。结论 持续暴露OVA联合SEB可成功诱导ECRS,该模型的建立为ECRS发病机制的研究提供基础。Objective The pathogenesis of eosinophilic chronic rhinosinusitis(ECRS) is not clear, so the establishment of an ideal animal model for related research has been paid more attention. At present, there is no report on the establishment of ECRS animal model in China. The aim of this study is to create a model of ECRS in mice using a common antigen(ovalbumin, OVA) plus a superantigen(Staphylococcal enterotoxin B, SEB). Methods Twenty-four female mice were randomly divided into four groups(six mice of each group):(1) the control group(the PBS group) was treated with phosphate buffered saline(PBS);(2) the SEB group was treated with SEB;(3) the OVA group was treated with OVA;(4) the OVA+SEB group was treated with OVA plus SEB. On day 103, nasal lavage fluid(NLF) was collected. The expression levels of interferon-γ(IFN-γ), interleukin-4(IL-4), IL-5 and IL-17 A from NLF were measured using enzyme-linked immunosorbent assay. Mice were sacrificed after obtaining NLF. The heads of mice were removed en bloc and fixed in formalin for histopathologic analysis. The histological changes of the sinonasal mucosa were examined by mean of hematoxylin, eosin staining, mainly to observe polyp-like lesions and eosinophilic infiltration. The expression of eosinophil cationic protein(ECP) in the sinonasal mucosa were assessed by immunohistochemical staining. Results The levels of IL-4 and IL-5 were markedly elevated in OVA and OVA+SEB groups compared with PBS and SEB groups(P<0.05). Meanwhile, mices in OVA+SEB group had higher IL-5 levels than that in OVA group(P<0.05). Polyp-like lesions and a large number of eosinophils were observed in the sinonasal mucosa in the OVA+SEB group. The numbers of polyp-like lesions and infiltrated eosinophils in the OVA+SEB group were statistically higher than those in the other three groups(P<0.05). Mice in OVA and OVA+SEB groups appeared to exhibit stronger positive signals for ECP in the sinonasal mucosa than mices in PBS and SEB groups, and the numbers of ECP-positive cells were significantly differ

关 键 词：嗜酸粒细胞性慢性鼻-鼻窦炎 疾病模型 动物 

分 类 号：R765[医药卫生—耳鼻咽喉科]