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目的探讨以重组粉尘螨Ⅱ类变应原(rDerf2)建立小鼠肺部变应性炎症模型的方法。方法将30只6～8周龄雌性SPF级BALB/c小鼠,随机分为PBS组(阴性对照组)、卵清蛋白(OVA)组(阳性对照组)和rDerf2组(实验组)。实验组分别于第0、7、14天用rDerf2对小鼠腹腔进行注射致敏;再于第21天开始,连续7d进行雾化激发;对照组分别用PBS和OVA代替。最后一次雾化激发后24h内处死动物,观察肺组织病理变化、支气管肺泡灌洗液(BLAF)中的白细胞计数及分类,用ELISA测定BLAF和脾细胞培养上清中白细胞介素2(IL-2)、IL-4、IL-17和IFN-γ的含量及血清中IgG1、IgE抗体水平变化。结果实验组和阳性对照组BALB/c小鼠肺组织呈现明显的炎症性病理改变;rDerf2组的BALF中白细胞总数〔(17.39±1.03)×106/L〕及嗜酸性粒细胞(EOS)〔(1.61±0.03)×106/L〕计数,均明显高于PBS组(P<0.01);BALF中IL-4〔(78.92±9.06)pg/ml〕、IL-17〔(201.63±31.26)pg/ml〕与PBS组比较呈明显的高水平表达(P<0.01);而IL-2〔(8.29±1.27)pg/ml〕和IFN-γ〔(51.04±15.85)pg/ml〕的含量则显著下降(P<0.01);上述指标在脾细胞培养上清中出现类似的变化。rDerf2组血清中IgE〔(37.63±6.57)IU/ml〕、IgG1〔(16.68±2.90)μg/ml〕的含量表现为Th2型反应增强趋势;但OVA组和rDerf2组间所有指标差异均无统计学意义(P>0.05)。结论用rDerf2腹腔注射致敏后再雾化激发的方式能够成功构建小鼠肺部变应性炎症模型。
Objective To investigate a method for establishing mouse lung allergic inflammation model by recombinant Dermatophagoides pteronyssum Ⅱ allergen (rDerf2). Methods Thirty female BALF / c mice of 6-8 weeks old were randomly divided into PBS group (negative control group), ovalbumin (OVA) group (positive control group) and rDerf2 group (experimental group). Experimental groups were injected with rDerf2 on day 0, 7 and 14, respectively, to sensitize the peritoneal cavity of mice. At the beginning of the 21st day, the mice were challenged by atomization for 7 days. The control group was replaced by PBS and OVA respectively. The animals were sacrificed 24h after the last challenge with atomization, and the pathological changes of lung tissue were observed. The white blood cell count and classification in bronchoalveolar lavage fluid (BLAF) were determined. The levels of interleukin 2 (IL- 2), IL-4, IL-17 and IFN-γ levels and serum IgG1, IgE antibody levels. Results The lung tissue of BALB / c mice in experimental group and positive control group showed obvious inflammatory pathological changes. The total number of leukocytes in BALF of rDerf2 group 〔(17.39 ± 1.03) × 106 / L〕 and eosinophil 〔(( (P <0.01). IL-4 〔(78.92 ± 9.06) pg / ml〕 and IL-17 〔(201.63 ± 31.26) pg / ml] was significantly higher than that in PBS group (P <0.01), while the levels of IL-2 〔(8.29 ± 1.27) pg / ml〕 and IFN-γ 〔(51.04 ± 15.85) pg / ml〕 (P <0.01). Similar changes occurred in the spleen cell culture supernatant. The serum levels of IgE (37.63 ± 6.57) IU / ml and IgG1 [(16.68 ± 2.90) μg / ml〕 in rDerf2 group showed an increasing trend of Th2-type reaction, but there was no statistical difference in all the indexes between OVA group and rDerf2 group Significance (P> 0.05). Conclusion The model of allergic inflammation in mice lungs can be successfully constructed by rDerf2 sensitized intraperitoneal injection and re-atomization.