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  

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

作　　者：沈小英[1,2] 朱清仙[2] 刘志刚[1] 李湘辉[1] 

机构地区：[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

出　　处：《中国寄生虫学与寄生虫病杂志》2009年第2期115-119,共5页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No.39860071,30571625,30613121);广东省科技计划重点专项(No.2003A3080502)~~

摘　　要：目的探讨重组德国小蠊变应原(rBlag2)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rBlag2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rBlag2,剂量为50mg/(只·次),共3次,每次间隔1周。A组以50ml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rBlag2,100mg/(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50mgrBlag2,连续7d。A组以PBS代替变应原滴鼻。于最后1次滴鼻后24h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rBlag2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rBlag2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rBlag2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。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）×10^5/ml]and eosinophils[（22.20±3.76）×10^5/ml] in BALF of group B significantly increased than those of group C[（14.30±4.95）×10^5/ml and（5.20±1.56）×10^5/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 inflammation of group C reduced considerably. Each test value of group C was substantially similar to that of group A. Conclusion The experiment s

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 

分 类 号：R757.9[医药卫生—皮肤病学与性病学]