【摘要】 目的:观察气道内T-bet基因转染对哮喘小鼠气道炎症的影响。方法:C57BL/6小鼠40只,随机分为4组,每组10只,分别为正常对照组(A组)、哮喘模型组(B组)、空质粒干预组(C组)和T-bet质粒干预组(D组)。卵白蛋白(OVA)抗原溶液腹腔注射致敏,滴鼻造模。正常对照组用生理盐水代替OVA,空质粒干预组和T-bet质粒干预组OVA激发48h前,分别经鼻滴入50μg空质粒和重组T-bet质粒。观察各组实验小鼠的肺组织炎症以及BALF中各类炎症细胞以及IL-4、IFN-γ水平的变化。结果:Western blotting检测发现,小鼠气道转染pcDNA3-T-bet质粒48h后肺组织T-bet蛋白表达显著增加。pcDNA3-T-bet质粒转染能较好抑制给药后48hOVA激发的哮喘小鼠气道炎症(包括炎症细胞浸润,上皮细胞损伤、黏液分泌、血管壁水肿及管腔缩窄);下调小鼠BALF中Th2因子IL-4并上调Th1因子IFN-γ水平。结论:气道内转染T-bet质粒能有效改善哮喘小鼠的气道炎症。更多还原

【Abstract】 AIM:To investigate the effect of T-bet plasmid gene transfer to airway on allergen induced airway inflammation in a murine asthmatic model. METHODS:A mouse asthma model was established by sensitization with ovalbumin (OVA). Forty C57BL/6 mice were divided into 4 groups (10 mice in each group):the normal control group (group A),the asthmatic model group (group B),the pcDNA3 plasmid group (group C),and the pcDNA3-T-bet group (group D). The animals in group B,C and D were sensitized and challenged with OVA. The animals in group A were applied with normal saline. pcDNA3 plasmid at dose of 50 μg was intranasally administered at 24 h before intranasal challenges to the mice in group C,and the 50 μg pcDNA3-T-bet plasmid for the mice in group D. Bronchial alveolar lavage fluid (BALF) was collected and lung tissues were resected at 48 h after OVA challenge for later assay. RESULTS:After administration with pcDNA3-T-bet plasmid,high level of T-bet expression at 48 h was detected in the lung tissue by Western blotting. In pcDNA3-T-bet treated asthmatic models,histological evaluation revealed the significant suppression of eosinophil peribronchial and perivascular infiltration,and reduction of epithelial damage. The numbers of eosinophils,neutrophils and lymphocytes in BALF from pcDNA3-T-bet treated mice were significantly reduced compared to those in asthmatic control group (P<0.05). The level of IL-4 in BALF was significantly decreased in pcDNA3-T-bet group compared to that in asthmatic control group (P<0.05),while the level of IFN-γ in BALF was significantly increased in pcDNA3-T-bet group. No significant change of inflammation cells and cytokines in pcDNA3 plasmid group and asthmatic control group was observed (P>0.05). CONCLUSION:Intranasal pcDNA3-T-bet plasmid transfer inhibits asthmatic airway inflammation in the murine asthmatic model,suggesting a new therapeutic strategy for allergic asthma.更多还原