重组粉尘螨Ⅱ类变应原致敏小鼠肺部变应性炎症模型的建立

中国媒介生物学及控制杂志 ›› 2012, Vol. 23 ›› Issue (3): 221-224.

重组粉尘螨Ⅱ类变应原致敏小鼠肺部变应性炎症模型的建立

周书林, 常维佳, 姜玉新, 陈文魁

皖南医学院医学寄生虫学教研室, 安徽芜湖 241002

收稿日期:2011-12-27 出版日期:2012-06-20 发布日期:2012-06-20
基金资助:
安徽省自然科学基金(11040606M214);安徽省教育厅自然科学研究项目(kj2009B152)

The model of pulmonary allergic inflammation in mice established with recombinant group 2 allergen of Dermatophagoides farinae

ZHOU Shu-lin, CHANG Wei-jia, JIANG Yu-xin, CHEN Wen-kui

Department of Medical Parasitology, Wannan Medical College, Wuhu 241002, Anhui Province, China

Received:2011-12-27 Online:2012-06-20 Published:2012-06-20
Supported by:
Supported by the Anhui Provincial Natural Science Fund(No. 11040606M214)and Natural Science Fund of Anhui Provincial Education Department(No. kj2009B152)

摘要/Abstract

摘要： 目的探讨以重组粉尘螨Ⅱ类变应原(rDer f 2)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rDer f 2组(实验组)。实验组分别于第0、7、14天用rDer f 2对小鼠腹腔进行注射致敏;再于第21天开始,连续7 d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24 h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;rDer f 2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。rDer f 2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rDer f 2组间所有指标差异均无统计学意义(P>0.05)。结论用rDer f 2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。

关键词: 粉尘螨, 重组粉尘螨Ⅱ类变应原, 致敏, 哮喘模型

Abstract: Objective To establish a model of pulmonary allergic inflammation with BALB/c mice. Methods BALB/c mice were divided into PBS group, OVA group and recombinant Der f 2 protein (rDer f 2) group. The mice in the test group were sensitized by intraperitoneal injection of rDer f 2 at day 0, 7 and 14, and exposed to aerosolized allergen for 7 consecutive days starting from the 21 days. PBS or OVA instead of rDer f 2 was used in the control groups. All mice were executed within 24 h after the final exposure to aerosolized allergen. Pathological changes of the lung, cell counts and classification were studied with IL-2, IL-4, IL-17 and IFN-γ levels in BALF detected by ELISA. Cells from spleen were cultured for 72 h,with IL-2, IL-4, IL-17 and IFN-γ in supernatant, and allergen-specific IgG1 and IgE in serum detected by ELISA. Results In the OVA and rDer f 2 groups, the total number of white blood cells [(17.39± 1.03)× 10⁶/L] and eosinophils [(1.61± 0.03)× 10⁶/L] in BALF were significantly higher in OVA and rDer f 2 groups than those in PBS group (P<0.01). The levels of IL-4 [(78.92±9.06) pg/ml] and IL-17 [(201.63±31.26) pg/ml] in BALF from OVA and rDer f 2 groups were significantly increased (P<0.01), compared with those in PBS group. On the contrary, levels of IL-2 [(8.29± 1.27) pg/ml] and IFN-γ [(51.04± 15.85) pg/ml] in BALF were reduced significantly (P<0.01). The similar trends were also detected in terms of the above cytokines in the supernatants of spleen cell cultures. Moreover, the levels of IgE [(37.63±6.57) IU/ml], IgG1 [(16.68±2.90) μg/ml] in OVA and rDer f 2 groups were also significantly increased in serum than those in PBS group (P<0.01). The phenomenon indicated that Th2-type immune response was enhanced and Th1-type immune response was reduced, with no statistically significant difference between OVA and rDer f 2 groups (P>0.05) observed. Conclusion A pulmonary allergic inflammation model of BALB/c mice can be established successfully by intraperitoneal injection followed by aerosolization with rDer f 2 protein.

Key words: Dermatophagoides farinae, Recombinant group 2 allergen, Allergy, Animal model of asthma

中图分类号:

R384.8

周书林, 常维佳, 姜玉新, 陈文魁. 重组粉尘螨Ⅱ类变应原致敏小鼠肺部变应性炎症模型的建立[J]. 中国媒介生物学及控制杂志, 2012, 23(3): 221-224.

ZHOU Shu-lin, CHANG Wei-jia, JIANG Yu-xin, CHEN Wen-kui. The model of pulmonary allergic inflammation in mice established with recombinant group 2 allergen of Dermatophagoides farinae[J]. Chines Journal of Vector Biology and Control, 2012, 23(3): 221-224.

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