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目的研究致敏原的剂量对哮喘模型免疫细胞活化及肺组织损伤的影响。方法以卵蛋白为致敏原,采用腹腔注射[高剂量组1 mg/(只·次),低剂量组0.1 mg/(只·次)]联合雾化吸入建立小鼠哮喘模型。ELISA分析血液中过敏相关免疫球蛋白IgE及细胞因子IL-4与IFN-γ的水平,免疫荧光及流式细胞术分析脾脏免疫细胞(包括T、B和DC细胞)的活化程度,HE染色分析肺组织的损伤情况。结果模型组小鼠血液中IgE及IL-4的水平明显高于对照组(P<0.05),且高剂量组高于低剂量组(P<0.05)。但IFN-γ的水平模型组均低于对照组(P<0.05),高、低剂量组间无差异(P>0.05)。模型组小鼠脾脏细胞膜型CD28、CD86、CD80、CD40和MHC-Ⅱ的表达均高于对照组(P<0.05),且高剂量组高于低剂量组(P<0.05)。显微镜下观察模型组小鼠肺组织出现炎性细胞浸润及支气管壁增厚等病理改变,高剂量组更为严重。结论哮喘引发的免疫细胞活化和肺组织病理损伤与致敏原的剂量呈正相关。
Objective To study the effect of allergen dose on immune cell activation and lung injury in asthmatic model. Methods The ovalbumin was used as the allergen to establish mouse asthma model by intraperitoneal injection [1 mg / (high dose), 0.1 mg / (low dose)) combined with inhalation. The levels of IgE, IL-4 and IFN-γ in blood were detected by ELISA. The activation of spleen immune cells (including T, B and DC cells) were analyzed by immunofluorescence and flow cytometry. HE staining Lung tissue damage. Results The levels of IgE and IL-4 in the blood of the model group were significantly higher than those of the control group (P <0.05), and were higher in the high-dose group than in the low-dose group (P <0.05). However, the level of IFN-γ in the model group was lower than that in the control group (P <0.05). There was no difference between the high and low dose groups (P> 0.05). The expression of membrane type CD28, CD86, CD80, CD40 and MHC-Ⅱ of spleen cells in model group were higher than those in control group (P <0.05), and were higher in high dose group than in low dose group (P <0.05). Under the microscope, pathological changes such as inflammatory cell infiltration and bronchial wall thickening were found in the lung tissue of the model group mice, and were more serious in the high-dose group. Conclusion The activation of immune cells induced by asthma and the pathological damage of lung tissue are positively correlated with the doses of allergens.