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目的探讨转录因子GATA-3在支气管哮喘小鼠发病机制中的作用及GATA-3圈套核苷酸治疗支气管哮喘的可行性。方法BALB/c小鼠32只,随机分为4组:正常对照组、哮喘模型组、GATA-3圈套核苷酸治疗组(ODN组)和突变圈套核苷酸治疗组(mODN组),每组8只。采用鸡卵白蛋白致敏方法建立支气管哮喘小鼠模型;通过呼吸道给药的方式,将GATA-3圈套核苷酸及突变GATA-3圈套核苷酸转染至哮喘小鼠肺部。应用免疫组织化学染色检测其GATA-3蛋白的表达,应用反转录聚合酶链反应检测小鼠肺组织IL-5、IL-13 mRNA表达;收集小鼠支气管肺泡灌洗液(BALF),进行细胞总数和嗜酸性粒细胞(EOS)计数,HE染色评价肺组织病理变化。结果哮喘模型组小鼠肺组织GATA-3蛋白表达水平、肺组织IL-5、13 mRNA表达水平、BALF细胞总数及EOS计数均明显高于正常对照组(Pa<0.01);ODN组小鼠肺组织IL-5、13 mRNA表达水平均明显低于哮喘模型组(Pa<0.01),BALF中细胞总数及EOS计数亦均较哮喘模型组显著减少(Pa<0.01),HE染色显示ODN组小鼠肺血管周围和支气管周围炎性反应明显减轻;而mODN组小鼠与哮喘模型组比较,上述指标均无明显变化。结论GATA-3高表达在小鼠哮喘模型效应阶段呼吸道炎性反应中具有重要作用,GATA-3圈套核苷酸通过特异性阻断GATA-3活性,有效抑制Th2型细胞因子的产生,从而减轻哮喘小鼠呼吸道炎性反应。GATA-3圈套核苷酸可作为一种新的方法治疗哮喘及其他以Th2型免疫反应为主的变应性疾病。
Objective To investigate the role of transcription factor GATA-3 in the pathogenesis of bronchial asthma and the feasibility of GATA-3 truncated nucleotides in the treatment of bronchial asthma. Methods Thirty-two BALB / c mice were randomly divided into 4 groups: normal control group, asthma model group, GATA-3 truncated nucleotides treatment group (ODN group), and mutated truncated nucleotide treatment group Group of eight. A mouse model of bronchial asthma was established by the method of chicken egg albumin sensitization. GATA-3 truncated nucleotides and mutant GATA-3 decoy nucleotides were transfused to the lungs of asthmatic mice by means of respiratory tract administration. The expression of GATA-3 protein was detected by immunohistochemical staining. The expression of IL-5 and IL-13 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). The bronchoalveolar lavage fluid (BALF) The total number of cells and eosinophils (EOS) count, HE staining lung pathological changes. Results The level of GATA-3 protein, the level of IL-5,13 mRNA, the total number of BALF and the number of EOS in the lung tissue of asthmatic model group were significantly higher than those in the normal control group (P <0.01) The expression of IL-5 and 13 mRNA in BALB / c mice was significantly lower than that in asthmatic model group (P <0.01), and the total number of cells and EOS count in BALF were significantly decreased compared with asthma model group (P <0.01) Inflammatory reaction around the perivascular and bronchioles was significantly reduced; while mODN group mice and asthma model group, the above indicators had no significant change. Conclusion GATA-3 overexpression plays an important role in airway inflammatory response in asthmatic model-induced stage mice. GATA-3 decoy nucleotides can effectively inhibit the production of Th2 cytokines by blocking GATA-3 activity, Asthmatic mice respiratory inflammatory response. GATA-3 decoy nucleotides can be used as a novel approach to treat asthma and other allergic diseases characterized by Th2-type immune responses.