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目的采用气管滴注方式进行甲苯二异氰酸酯(toluene-diisocyanate,TDI)激发给药,改进TDI哮喘小鼠模型建立方法,并对模型的建立进行评价。方法将20只SPF级健康雌性BALB/c小鼠随机分为TDI组和溶剂对照组,每组10只。TDI组小鼠第1、8天于耳背分别涂抹20μl 0.3%TDI致敏,第15天采用气管滴注法给予20μl 0.01%TDI进行激发,溶剂对照组给予等量溶剂(丙酮与橄榄油混合物)。激发结束后,观察各组小鼠的行为学改变。致敏实验后每组取6只小鼠收集支气管肺泡灌洗液(BALF)并进行炎细胞计数分类,以ELISA法检测BALF上清中白细胞介素-4(IL-4)、γ-干扰素(IFN-γ)含量;以HE染色观察小鼠肺组织病理变化。结果激发结束后,TDI组小鼠均出现呼吸急促、点头呼吸、弓背、前肢缩抬等行为学改变,溶剂对照组未出现上述症状。TDI组小鼠BALF中白细胞总数、嗜中性粒细胞、淋巴细胞、单核细胞、嗜酸性粒细胞、嗜碱性粒细胞数目均高于溶剂对照组,且嗜中性粒细胞、嗜酸性粒细胞百分比高于溶剂对照组,BALF中细胞因子IL-4、IFN-γ含量高于溶剂对照组,差异均有统计学意义(P<0.05)。HE染色可见TDI组小鼠肺组织内支气管管壁破坏明显,管腔增厚,支气管管腔内及肺泡腔内有大量的炎细胞浸润,而溶剂对照组未见明显病理变化。结论采用气管滴注方式进行TDI激发给药,可成功建立TDI哮喘小鼠模型。
OBJECTIVE: To establish a method of TDI asthma mouse model by using tracheal instillation, and to establish the model by using toluene-diisocyanate (TDI). Methods Twenty SPF healthy female BALB / c mice were randomly divided into TDI group and solvent control group, with 10 mice in each group. TDI mice were sensitized with 20μl 0.3% TDI on the dorsum of their ears on the 1st and the 8th day, respectively, and 20μl of 0.01% TDI was administered by tracheal instillation on the 15th day. The solvent control group was given equal amount of solvent (mixture of acetone and olive oil) . After the stimulation, the behavioral changes of the mice in each group were observed. After sensitization, BALB / c mice were collected from 6 mice in each group and their inflammatory cell count was collected. The levels of interleukin-4 (IL-4), interferon-γ (IFN-γ) content; the pathological changes of lung tissue were observed by HE staining. Results After the stimulation, the mice in TDI group developed the behavior changes such as shortness of breath, nostril breathing, dorsal bowel, forelimb contraction and so on. The solvent control group did not have the above symptoms. The number of leukocytes, neutrophils, lymphocytes, monocytes, eosinophils and basophils in BALF of TDI group were all higher than that of solvent control group, and neutrophils, eosinophils The percentage of cells in the BALF was higher than that in the solvent control group. The content of IL-4 and IFN-γ in the BALF was higher than that in the solvent control group (P <0.05). In the TDI group, the damage of the bronchial wall in the lung tissue of the mice in the TDI group was obvious, the lumen thickening, the bronchial lumen and the alveolar cavity had a large number of infiltration of inflammatory cells, while the solvent control group showed no obvious pathological changes. Conclusions The TDI asthmatic mouse model can be successfully established by intratracheal instillation of TDI.