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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 investigate the effect of intranasal IFNγ gene transfection on airway inflammation in bronchial asthma (asthma) mice. Methods Forty healthy 6-week-old SPF C57BL / 6 mice were divided into 4 groups according to random number table. Normal control group (group A), asthma model group (group B), model empty plasmid intervention group (group C), model IFNγ plasmid intervention group (group D), 10 rats in each group. Groups B, C and D were sensitized intraperitoneally with 0.1 ml ovalbumin (OVA) 0.1 ml intraperitoneally, and asthmatic model was established by intranasal challenge with 1% OVA 50μl intranasally. In group A, 0.1% OVA 0.1ml And 1% OVA 50μl. Groups C and D were intranasally instilled with 50μl of empty plasmid pcDNA3.1 () and Lipofentamine2000 mixture or 50μl of recombinant IFNγ plasmid and Lipofentamine2000 mixture respectively. The cellular components, interleukin 4 (IL4), IL5, IFNγ and lung histopathological changes in bronchoalveolar lavage fluid (BALF) of mice in each group were observed. Results The total number of inflammatory cells, eosinophils, neutrophils and lymphocytes in BALF of group B were (0.102 ± 0.020) × 109 / L, (0.0193 ± 0.0047) × 109 / L, (0.0107 ± 0.0039) × 109 / L, (0.0255 ± 0.0042) × 109 / L in group A, (0.082 ± 0.012) × 109 / L in group A (P <0.05). The total number of inflammatory cells, EOS, neutrophils and lymphocytes in BALF in group D were significantly higher than that in group A The cell counts were (0.086 ± 0.016) × 109 / L, (0.0116 ± 0.0031) × 109 / L, (0.0062 ± 0.0018) × 109 / L and (0.0182 ± 0.0041) × 109 / L respectively Statistical significance (P <0.05). The levels of IL4, IL5 and IFNγ in BALF in group B were (39.2 ± 5.1) pg / ml, (83.7 ± 4.7) pg / ml and (15.7 ± 2.7) pg / ml, respectively) (13.3 ± 1.9) pg / ml, (12.1 ± 2.3) pg / ml and (31.8 ± 7.9) pg / ml respectively). There was significant difference between groups A and B The levels of IL-4, IL-5 and IFN-γ were (16.4 ± 3.2) pg / ml, (26.3 ± 3.4) pg / ml and 65.4 ± 10.4 pg / ml, respectively. 0.05). There was no obvious inflammatory changes in the airway of group A mice, the infiltration of inflammatory cells in the bronchus, submucosal and surrounding lung tissues of mice in groups B and C, and obvious edema of blood vessel wall. The airway inflammation in group D was significantly reduced. Conclusions Transfection of interferon plasmid in the airway can effectively improve the abnormality of cytokines IL4, IL5 and IFNγ in asthmatic mice, meanwhile inhibit EOS, neutrophil number and recruitment of lymphocytes in lungs and inflammation of lung tissue.