中性粒细胞性支气管哮喘小鼠模型建立的探索

目的利用不同剂量脂多糖（lipopolysaccharides,LPS）干预卵清蛋白（ovalbumin,OVA）致敏和激发的小鼠,探索中性粒细胞性哮喘（neutrophilic asthma,NA）小鼠模型的建立。方法 80只BALB/c小鼠随机分为正常对照组（A组）、肺损伤对照组（B组：B1～B4）、嗜酸细胞性哮喘模型对照组（C组）、NA模型探索组（D组：D1～D4）。分致敏和激发两阶段建模,致敏阶段：A组PBS腹腔注射及PBS滴鼻,B组PBS腹腔注射及LPS滴鼻,C组OVA腹腔注射,D组OVA腹腔注射及LPS滴鼻;激发阶段：A、B组生理盐水雾化,C、D组5%OVA雾化。通过观察小鼠雾化时的症状、肺组织病理改变,检测支气管肺泡灌洗液（bronchoalveolar lavage fluid,BALF）细胞总数及分类计数和血清OVA-sIgE等评价NA小鼠模型建立。结果①D3组小鼠出现类似C组小鼠呼吸急促、大小便失禁等表现。②D3组BALF炎症细胞总数较A组显著升高（P〈0.01）;D3组中性粒细胞（neutrophil,NEU）百分比（NEU%）较A、C、D1、D2组显著升高（P值均〈0.01）,嗜酸粒细胞（eosinophil,EOS）百分比（EOS%）较A组显著升高而较C组显著低下（P值均〈0.01）。③D3组支气管管壁及肺泡间隔增厚变形,部分断裂,气道黏膜下除了EOS浸润外还存在明显NEU浸润。④D3组血清OVA-sIgE水平显著高于A组而低于C组（P值均〈0.05）。⑤D3组IL-4水平显著高于A组（P〈0.05）,与C组无显著差异（P〉0.05）;D3组IFN-γ水平显著低于A组（P〈0.05）,与C组无显著差异（P〉0.05）。结论 10μg LPS滴鼻吸入＋50μg OVA腹腔注射三次与5%OVA连续激发两周即D3组可成功建立NA小鼠模型。

Exploration of a mice model of neutrophilic asthma

Objective To establish a mice model of neutrophilic asthma with different doses of lipopolysaccharide on a mice model of allergic asthma.Methods 80 BALB/c mice were randomly divided into four groups:control group(group A),lung injury groups(group B:B1-B4),eosinophilic asthma group(group C),neutrophilic asthma groups(group D:D1-D4).These were divided into sensitized and stimulated phase,sensitized phase:group A:intra-peritoneal injection with PBS and nasal drip with PBS,group B:intra-peritoneal injection with PBS and nasal drip with LPS(lipopolysaccharides),group C:intra-peritoneal injection with ovalbumin(OVA),stimulated phase:group A and B:atomization with saline,group C and D:atomization with 5% OVA.Physiologic responses to atomization and change of lung tissue were observed.Total cell counts and classification in bronchoalveolar lavage fluid(BALF) were measured.OVA-specific IgE levels in serum were detected for evaluating the establishment of the model.Results ①The mice of group D3 presented fast breathing,urinary and fecal incontinence.②The total cell counts of group D3 in BALF were significantly increased compared with group A(P<0.01).The percentage of neutrophils of group D3 was significantly increased compared with group A,C,D1 and D2(all P<0.01).The percentage of eosinophils of group D3 was significantly increased compared with group A but decreased compared with group C(both P<0.01).③The airway wall thickening,partial ruptured,and more neutrophils infiltration in the airway mucosa and submucosa could be seen in lung pathology of group D3.④Levels of OVA-sIgE in serum of group D3 were significantly higher than group A but lower than group C(both P<0.05).⑤Levels of IL-4 in BALF of group D3 was significantly higher than group A but IFN-γ was lower than group A(both P<0.05),both are no significant differences with group C(both P>0.05).Conclusions A mice model of NA could be successfully established with intranasal applications of 10 μg LPS+intraperitoneal injection of 50 μg OVA sensibilized three times and with 5% OVA aerosol inhalation challenged for two weeks.