哮喘小鼠下呼吸道流感嗜血杆菌定植的研究

目的：建立哮喘合并下呼吸道流感嗜血杆菌定植小鼠模型,并进行评价。方法：将80只小鼠按随机数字表法分为正常组、正常注菌（流感嗜血杆菌）组 、哮喘组 、哮喘注菌（流感嗜血杆菌）组。哮喘组、哮喘注菌组用卵清蛋白（ovalbumin, OVA）致敏和激发小鼠制作哮喘模型,正常注菌组、哮喘注菌组小鼠气管内滴注琼脂包被的流感嗜血杆菌,每组各取一半小鼠分别在气道滴注之后的第7天和第21天处死,取肺组织进行病理观察,行气管细菌培养,测定肺泡灌洗液（broncheoalveolar lavage fluid,BALF）的细胞计数,检测血清和BALF中肿瘤坏死因子α（tumor necrosis factor α,TNF-α）水平。结果：与正常注菌组及哮喘组（未注菌）相比,哮喘注菌组小鼠有明显的哮喘和感染表现;病理学结果显示其气道管壁增厚、黏膜水肿,管腔内有黏液分泌,且有大量炎症细胞浸润;气道形态学参数分析显示,其气管壁及气管平滑肌增厚;BALF中白细胞总数和中性粒细胞计数增多;血清和BALF中TNF-α水平升高;注菌后第7天和第21天气管细菌培养可见流感嗜血杆菌。结论：用OVA制备哮喘动物模型,并在气道内注入琼脂包被的流感嗜血杆菌,可以建立哮喘合并呼吸道流感嗜血杆菌定植的动物模型,造成肺部气道重塑和炎症反应。

An asthma mice model with airway colonized by Haemophilus influenzae

Objective: To establish an asthma mice model with airway colonized by Haemophilus influenzae. Method: Eighty BALB/C mice were grouped into 4 groups: normal control, Haemophilus influenza colonization, asthma control, asthma with Haemophilus influenza colonization. Mice asthma was induced by sensitizing and challenging with ovalbumin（OVA）, and airway Haemophilus influenza colonization was established by intratracheal instillation of agar-coated Haemophilus influenza. Half of the mice in each group were sacrificed at 7th day, and the other half at 21 th day. Lung tissue was obtained for histopathologic examination and bacterial culture; serum and bronchoaveolar fluid (BALF) TNF-α levels were detected, and cell count in BALF was determined. Results: Compared with normal control and asthma control groups, asthma with Haemophilus influenza colonization group showed obviously asthma and infection symptoms, with pathological characteristics including obvious airway thickening, mucosa edema, mucus secretion and inflammatory cells infiltration in lung tissue. WBC and neutrophils count in BALF increased significantly and level of TNF-α in serum and BALF were significantly higher in asthma with Haemophilus influenzae colonization group; Haemophilus influenzae was found in bacteria culture. Conclusions: Asthma mice model with airways colonized by Haemophilus influenzae could be established by intratracheal instillation of agar-coated Haemophilus influenzae in asthma mice induced by OVA sensitizing and challenging.