德国小蠊重组变应原（rBla g 2）治疗过敏性哮喘小鼠的实验研究

中国寄生虫学与寄生虫病杂志 ›› 2009, Vol. 27 ›› Issue (2): 5-119.

德国小蠊重组变应原（rBla g 2）治疗过敏性哮喘小鼠的实验研究

沈小英^1，2，朱清仙²，刘志刚^{1 *}，李湘辉¹

1 深圳大学医学院过敏反应与免疫学研究所，深圳 518060；2 南昌大学基础医学院，南昌 330006

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-04-30 发布日期:2009-04-30

Experimental Study on Recombinant Bla g 2（rBla g 2） in the Treatment of Allergic Asthma in Mice

SHEN Xiao-ying^1，2，ZHU Qing-xian²，LIU Zhi-gang^{1 *}，LI Xiang-hui¹

1 Institute of Allergy and Immunology，School of Medicine，Shenzhen University，Shenzhen 518060，China；2 College of Basic Medicine，Nanchang University，Nanchang 330006，China

Received:1900-01-01 Revised:1900-01-01 Online:2009-04-30 Published:2009-04-30

摘要/Abstract

摘要： 目的探讨重组德国小蠊变应原（rBla g 2）治疗过敏性哮喘小鼠的效果及机制。方法 18只BALB/c小鼠随机均分为阴性对照组（A组）、哮喘模型组（B组）和重组蛋白rBla g 2治疗组（C组）。B、C两组分别腹腔注射经Al（OH）3佐剂乳化的重组蛋白rBla g 2，剂量为50 mg/（只?次），共3次，每次间隔1周。A组以50 ml生理盐水代替变应原。末次致敏后2周，进行免疫治疗，C组腹腔注射rBla g 2，100 mg/（只?次），每两天1次，连续8次。A、B两组以PBS代替变应原。末次治疗后1周，将小鼠麻醉，B、C两组每鼠每天滴鼻50 mg rBla g 2，连续7 d。A组以PBS代替变应原滴鼻。于最后1次滴鼻后24 h检测各项指标：各组小鼠气道高反应性，支气管肺泡灌洗液（BALF）中细胞总数和细胞种类以及血清中rBla g 2抗原特异性IgE和IgG2a的变化；HE染色观察小鼠肺组织的改变，免疫组化检测肺嗜酸粒细胞（EOS）B淋巴细胞瘤/白血病-2（Bcl-2）的表达。结果与B组相比，C组气道高反应检测中扩大间隙（Penh）值下降（P＜0.05）；血清中rBla g 2特异性IgE降低，IgG2a增加（P＜0.01）；C组EOS细胞阳性数量明显减少，且Bcl-2蛋白阳性表达较弱。B组BALF的细胞总数和EOS数分别为（24.60±15.08）×105个/ml和（22.20±3.76）×105个/ml，而C组细胞总数［（14.30±4.95）×105个/ml］和EOS数［（5.20±1.56）×105个/ml］显著减少（P＜0.01）。B组肺部气管周围炎性细胞聚集，肺组织上皮损伤，组织水肿，而C组肺部变应性炎症明显减轻。与A组相比，C组各项检测指标接近A组。结论 rBla g 2对小鼠过敏性哮喘有治疗作用，可能是EOS细胞凋亡在其中起重要作用。

关键词: 德国小蠊, 重组变应原, 哮喘, 嗜酸粒细胞, B淋巴细胞瘤/白血病-2

Abstract: Objective To study the therapeutic effect on murine allergic asthma with recombinant Bla g 2 （rBla g 2）allergen and its possible mechanism. Methods Eighteen BALB/C mice were randomly divided into three groups: normal control group（group A）, asthma model group（group B）, and recombinant protein rBla g 2 treatment group（group C）. Mice in groups B and C were subcutaneously immunized weekly with rBla g 2（50 mg）formulated in Al（OH）3 adjuvant for three weeks. Group A received only adjuvant emulsified with normal saline. Two weeks after the last inoculation, mice in group C were administered each with rBla g 2（100 mg）/dose, and groups A and B were given PBS. All the mice received eight doses at 2-day intervals. One week after the last immunotherapy, mice in groups B and C were intranasally challenged with 50 mg rBla g 2 daily for seven days, while mice in group A received PBS. Twenty-four hours after the challenge, the following items were examined: airway hyperresponsiveness of mice, total cellular score and cell classification in bronchoalveolar lavage fluid（BALF）, level of rBla g 2-specific IgE and IgG2a in serum, lung inflammation by HE stain, and Bcl-2 expression of eosinophils of lung by immunohistochemistry. Results Compared with group B, group C showed a decreased Penh value of airway hyperresponsiveness（P＜0.05）, reduced serum rBla g 2-specific IgE but increased IgG2a（P＜0.01）, and reduced Bcl-2 expression of eosinophils. Total cells［（24.60±15.08）×105/ml］and eosinophils［（22.20±3.76）×105/ml］ in BALF of group B significantly increased than those of group C［（14.30±4.95）×105/ml and（5.20±1.56）×105/ml，respectively］（P＜0.01）. The interstitial space surrounding the airway lumen was characterized by a denseｌｙ mixed cellular infiltrate, tissue edema and epithelium tissue damage in group B, while lung inflamma-tion of group C reduced considerably. Each test value of group C was substantially similar to that of group A. Conclu-sion The experiment shows proper immunotherapeutic efficacy of rBla g 2 in murine allergic asthma, which may possi-bly related to the apoptosis of eosinophils.

Key words: Blatella germanica, Recombinant Bla g 2（rBla g 2）, Asthma, Eosinophil, Bcl-2

沈小英;朱清仙;刘志刚;李湘辉. 德国小蠊重组变应原（rBla g 2）治疗过敏性哮喘小鼠的实验研究[J]. 中国寄生虫学与寄生虫病杂志, 2009, 27(2): 5-119.

SHENXiao-ying;ZHUQing-xian;LIUZhi-gang*;LIXiang-hui. Experimental Study on Recombinant Bla g 2（rBla g 2） in the Treatment of Allergic Asthma in Mice[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2009, 27(2): 5-119.

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