Jia Aijun, Wang Yueling, Sun Wenjin, Xiao Bing, Mu Lin, Wei Yan, Xu Li, Peng Cong, Zhang Dongshan, Shen Huahao, Xiang Xudong
Department of Respiratory Medicine, The Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, P.R. China.
Department of Anesthesiology, Xiangya Hospital of Central South University, Changsha, Hunan 410008, P.R. China.
Exp Ther Med. 2017 Sep;14(3):2126-2134. doi: 10.3892/etm.2017.4776. Epub 2017 Jul 11.
Neutrophilic asthma (NA) is associated with a severe disease course and poor response to corticosteroids. The present study aimed to compare the effects of various concentrations of house dust mite (HDM) allergens, ovalbumin (OVA), the major egg allergen, and lipopolysaccharide (LPS) in combination on the onset of severe NA. Female C57BL/6 mice were grouped according to a random number table and intranasally sensitized with HDM/LPS/OVA extracts on days 0, 1 and 2 of the study. In group 1, mice received 50 µg HDM + 50 µg OVA + 15 µg LPS, mice in group 2 received 50 µg HDM + 100 µg OVA + 15 µg LPS, mice in group 3 received 100 µg HDM + 50 µg OVA + 15 µg LPS and those in group 4 received 100 µg HDM + 100 µg OVA + 15 µg LPS, while mice in the control group received saline only. The mice were then challenged by OVA solution with atomized excitation on days 14, 15, 18, 19 and 20 for 30 min each. Ethology, airway hyperresponsiveness (AHR), immune cell distributions in bronchoalveolar lavage fluid (BALF), and specific cytokines interleukin 17A (IL-17A) and IL-4 in serum were assessed. Histological examination of inflammation by hematoxylin and eosin staining and immunohistochemical assessment of neutrophils (NEU), eosinophils (EOS), IL-17A and IL-4 were also performed. Compared with the control group, the HDM/OVA/LPS-sensitized groups 1-4 had markedly increased BALF cells, serum interleukin IL-17A and IL-4, inflammatory cell infiltration, EOS as well as IL-17A and IL-4 by immunohistochemical staining (all P<0.05). Among the four HDM/OVA/LPS-sensitized groups, mice of group 4 had higher AHR, a significantly higher total cell number, NEU and EOS in BALF as well as significantly higher NEU and NEU/EOS ratios according to immunohistochemical staining when compared to groups 1-3 (P<0.05 for all). In conclusion, sensitization with 100 µg HDM + 100 µg OVA + 15 µg LPS successfully established a severe asthma model with a predominantly neutrophilic inflammatory phenotype.
嗜中性粒细胞性哮喘（NA）与严重的病程和对皮质类固醇的不良反应相关。本研究旨在比较不同浓度的屋尘螨（HDM）过敏原、卵清蛋白（OVA，主要的鸡蛋过敏原）和脂多糖（LPS）联合使用对严重NA发病的影响。将雌性C57BL/6小鼠根据随机数字表分组，并在研究的第0、1和2天经鼻用HDM/LPS/OVA提取物进行致敏。在第1组中，小鼠接受50μg HDM + 50μg OVA + 15μg LPS，第2组小鼠接受50μg HDM + 100μg OVA + 15μg LPS，第3组小鼠接受100μg HDM + 50μg OVA + 15μg LPS，第4组小鼠接受100μg HDM + 100μg OVA + 15μg LPS，而对照组小鼠仅接受生理盐水。然后在第14、15、18、19和20天用OVA溶液雾化激发对小鼠进行攻击，每次30分钟。评估行为学、气道高反应性（AHR）、支气管肺泡灌洗液（BALF）中的免疫细胞分布以及血清中的特异性细胞因子白细胞介素17A（IL-17A）和IL-4。还通过苏木精和伊红染色对炎症进行组织学检查，并对中性粒细胞（NEU）、嗜酸性粒细胞（EOS）、IL-17A和IL-4进行免疫组织化学评估。与对照组相比，HDM/OVA/LPS致敏的第1 - 4组BALF细胞、血清白细胞介素IL-17A和IL-4、炎症细胞浸润、EOS以及免疫组织化学染色显示的IL-17A和IL-4均明显增加（所有P<0.05）。在四个HDM/OVA/LPS致敏组中，与第1 - 3组相比，第4组小鼠的AHR更高，BALF中的总细胞数、NEU和EOS显著更高，并且根据免疫组织化学染色，NEU和NEU/EOS比值也显著更高（所有P<0.05）。总之，用100μg HDM + 100μg OVA + 15μg LPS致敏成功建立了以嗜中性粒细胞为主的炎症表型的严重哮喘模型。
