摘要
目的观察气道内γ干扰素(IFNγ)基因转染对支气管哮喘(简称哮喘)小鼠气道炎症的影响。方法健康6周龄SPF级C57BL/6小鼠40只,按随机数字表法分为4组。正常对照组(A组)、哮喘模型组(B组)、模型空质粒干预组(C组)、模型IFNγ质粒干预组(D组),每组10只。B组、C组及D组以0.1%卵白蛋白(OVA)0.1ml腹腔注射致敏,以1%的OVA50μl滴鼻激发建立哮喘模型;A组用等量生理盐水分别代替0.1%的OVA0.1ml和1%OVA50μl;C组和D组分别经鼻滴入空质粒pcDNA3.1()和Lipofentamine2000混合液50μl或重组IFNγ质粒和Lipofentamine2000混合液50μl。观察各组小鼠支气管肺泡灌洗液(BALF)中的细胞成分、白细胞介素4(IL4)、IL5、IFNγ和肺组织病理学变化。结果B组小鼠BALF中炎性细胞总数、嗜酸粒细胞(EOS)、中性粒细胞和淋巴细胞计数分别为(0.102±0.020)×109/L、(0.0193±0.0047)×109/L、(0.0107±0.0039)×109/L、(0.0255±0.0042)×109/L,A组分别为(0.082±0.012)×109/L、(0.0041±0.0009)×109/L、(0.0051±0.0016)×109/L、(0.0201±0.0019)×109/L,A、B两组比较差异有统计学意义(P<0.05);D组小鼠BALF中炎性细胞总数、EOS、中性粒细胞和淋巴细胞计数分别为(0.086±0.016)×109/L、(0.0116±0.0031)×109/L、(0.0062±0.0018)×109/L、(0.0182±0.0041)×109/L,与B组比较差异有统计学意义(P<0.05)。B组小鼠BALF中IL4、IL5、IFNγ水平分别为[(39.2±5.1)pg/ml、(83.7±4.7)pg/ml、(15.7±2.7)pg/ml],A组小鼠分别为[(13.3±1.9)pg/ml、(12.1±2.3)pg/ml、(31.8±7.9)pg/ml],A、B两组比较差异有统计学意义(P<0.05);D组小鼠BALF中IL4、IL5、IFNγ水平分别为[(16.4±3.2)pg/ml、(26.3±3.4)pg/ml、(65.4±10.4)pg/ml],与B组比较差异有统计学意义(P<0.05)。A组小鼠气道无明显炎症变化,B和C组小鼠小支气管、血管黏膜下和周围肺组织有明显的炎症细胞浸润,血管壁明显水肿;D组小鼠气道炎症明显减轻。结论气道内转染干扰素质粒能有效改善哮喘小鼠细胞因子IL4、IL5和IFNγ异常,同时对EOS、中性粒细胞数和淋巴细胞肺内募集、肺组织炎症改变有一定抑制作用。
Objective To study the effect of airway gamma interferon （IFN-γ） plasmid gene transfer on airway inflammation in asthmatic mice. Methods Forty C57BL/6 mice were randomly divided into four groups： a control group（group A）, an asthmatic group（group B）, a plasmid group （group C） and an IFN-γ plasmid group（group D）, 10 mice in each group. Except group A, other groups were sensitized with 0. 1 ml 0. 1% ovalbumin（OVA） by a combination of intraperitoneal injection and repeated 50 μl 1% OVA intranasal challenges to establish the mouse asthma model. In group A, normal saline of the equal volume was given instead of 0. 1% OVA 0. 1 ml and 1% OVA 50 μl. Group C was intranasally administered 50 μl mixture of plasmid pcDNA3. 1 （-） and Lipofentamine 2000, while 50 μl mixture of IFN-γ, plasmid and Lipofentamine 2000 was administered for the mice of group D. Bronchoalveolar lavage fluid （BALF） cell count and differential were studied. Interleukin-4 （ IL-4 ）, interleukin-5 （ IL-5 ） and IFN-γ in BALF were determined. Pathologic changes in lung tissues were observed. Results The differences were significant （ P 〈 0. 05） when the numbers of inflammation cells, eosinophils, neutrophils and lymphocytes in BALF of group B [（0. 102 ±0.020） x 10^9/L, （0.019 3 ±0.004 7） x 10^9/L, （0.010 7 ±0.003 9） x 10^9/L, （0. 025 5 ± 0. 004 2） x 10^9/L, respectively ] were compared with those of group A [ （0. 082 ± 0. 012） x 10^9/L,（0.004 1 ±0.000 9） x 10^9/L, （0.005 1 ±0.001 6） x 10^9/L, （0.020 1 ±0.001 9） x 10^9/L, respectively]. The numbers of inflammation cells, eosinophils, neutrophils and lymphocytes in BALF of group D [（0.086 ±0.016） x 10^9/L, （0.011 6±0.003 1） x 10^9/L, （0.006 2 ± 0.001 8） x 10^9/L, （0. 018 2 ± 0. 004 1 ） x 10^9/L, respectively] were also significantly different （P 〈 0. 05 ） as compared with those of group B. The IL--4, IL-5 and IFN-γ levels in BALF of group B [ （39. 2 ± 5. 1 ） pg/ml, （ 83.7± 4. 7 ） pg/ml, （ 15.7 ± 2. 7 ） pg/ml, respectively ] were significantly different （ P 〈 0. 05 ） as compared with those of group A [ （ 13.3 ± 1.9） pg/ml, （ 12. 1 ± 2. 3 ） pg/ml, （ 31.8 ± 7.9） pg/ml, respectively ]. The IL-4, IL-5 and IFN-γ levels of group D[ （ 16. 4 ± 3. 2） pg/ml, （26. 3 ± 3.4） pg/ml, （65.4 ± 10. 4） pg/ml ] were also different （ P 〈 0. 05 ） from those of group B. Lung inflammation was examined in HE stained sections. There was no obvious infiltration of inflammatory cells in the airways of group A. However, there were a great number of inflammatory cells in the interstitial and peribronchovascular regions of group B and group C. Group D exhibited reduced epithelial damage and less infiltration of mononuclear cells and polymorphs in the interstitial and peribronchovascular regions, as compared with group B or group C. Conclusions These findings suggest that transtracheal IFN-γ gene transfer is effective in modulating the imbalance of Th1/Th2 in BALF and inhibiting airway inflammation of asthmatic mice. The result provides experimental data for developing a novel therapeutic approach to asthma.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
2005年第8期530-533,共4页
Chinese Journal of Tuberculosis and Respiratory Diseases
基金
广东省自然科学基金重点攻关资助项目(A000099036)
