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观察呼吸道局部由逆转录病毒介导的IL-4RA(IL-4受体拮抗剂)基因转染对哮喘模型过敏性炎症以及气道高反应性的干预作用.BALB/c小鼠被随机分为5组(每个实验每组10只):空白对照组、哮喘模型组、激素治疗组、目的基因(1×104CFU/mLpLNC-IL-4RA)干预组和空载体(1×104CFU/mLpLNC-laz)干预组.各组在最后一次激发后24h进行气道高反应性的检测,之后放血杀死小鼠,检测血中嗜酸性粒细胞计数以及OVA(鸡卵白蛋白)特异性IgE,用ELISA方法检测BALF(支气管肺泡灌洗液)中的各种细胞因子水平并对BALF中各种炎症细胞计数并分类,肺组织切片HE染色.用免疫组化的方法检测STAT6(信息传递与转录活化因子,signal transducers and activators of transcription 6)的表达情况.转染有IL-4RA基因的哮喘模型鼠肺组织进行PCR扩增观察IL-4RA基因在宿主基因组的整合情况.结果显示,IL-4RA基因转染明显减轻了哮喘模型的气道高反应性,另外哮喘模型鼠BALF和血中有大量嗜酸细胞,肺组织病理切片可见大量炎症细胞浸润,哮喘模型的这些改变在激素和pLNC-IL-4RA干预后明显减轻,统计学分析有显著差异(P<0.01).激素和pLNC-IL-4RA干预后同时也明显减轻了哮喘模型组BALF中IL-5,IL-13的水平,差异有显著性(P<0.01),哮喘模型组STAT6的表达较空白对照组明显增多,而在激素和pLNC-IL-4RA干预后表达明显减少.结果提示,通过逆转录病毒转染的IL-4RA的基因高表达明显减轻了哮喘模型的气道高反应性、成功地阻止在哮喘模型鼠过敏性炎症以及气道炎症细胞浸润,调整了免疫失衡,因此基因治疗可能会成为治疗慢性哮喘气道炎症和哮喘症状的极有潜力的药物.
To investigate the intervention of local retrovirus-mediated transfection of IL-4RA (IL-4 receptor antagonist) gene in respiratory tract on allergic inflammation and airway hyperresponsiveness in asthmatic model.BALB / c mice were randomly divided into (10 × 10 4 CFU / mL pLNC-laz) groups (n = 10 in each experiment): the blank control group, the asthma model group, the hormone treatment group, the target gene ) Intervention group.The airway hyperresponsiveness was detected 24h after the last challenge in each group, after which the mice were bled to kill the mice, the blood eosinophil count and OVA (chicken egg albumin) -specific IgE were detected by ELISA The levels of cytokines in BALF (bronchoalveolar lavage fluid) were detected, and various inflammatory cells in BALF were counted and classified, and the lung tissue sections were stained with HE.The expression of STAT6 (signal transducers and activators of transcription, signal transducers and activators of transcription 6) .The IL-4RA gene was transfected into the lung tissue of mice with asthma to amplify the IL-4RA gene in the host genome.The results showed that IL-4RA gene transfection Significantly reduced the airway height of the asthma model In addition, a large number of eosinophils were found in the BALF and blood of the asthmatic model rats. A large number of infiltrating inflammatory cells could be seen in the pathological sections of the lungs. These changes in the asthmatic model were significantly reduced after the intervention of the hormone and pLNC-IL-4RA. Statistical analysis was significant (P <0.01) .After the intervention of hormone and pLNC-IL-4RA, the level of IL-5 and IL-13 in BALF of asthmatic model group was also significantly reduced (P <0.01) The expression of IL-4RA gene was significantly increased compared with that of the control group, but significantly decreased after the intervention of hormone and pLNC-IL-4RA.The results indicated that the high gene expression of IL-4RA transfected by retrovirus significantly reduced the airway height Reactivity, which successfully blocks allergic inflammation and airway inflammatory cell infiltration in the asthmatic model mice and adjusts for immune imbalance, gene therapy may be a potential drug for the treatment of airway inflammation and asthma symptoms in chronic asthma.