核因子κB顺式诱饵元件对哮喘大鼠模型治疗作用的观察

The therapeutic efficacy of nuclear factor κB decoy cis element in animal model of asthma

目的 探讨核因子κB(NF κB)顺式诱饵元件治疗支气管哮喘的可行性。方法 通过从气道局部给药的方式 ,将NF κB顺式诱饵寡核苷酸 (N ODN)或无序诱饵ODN(S ODN)转染至哮喘大鼠肺部。运用原位组织杂交、反转录PCR、免疫组织化学、酶联免疫吸附试验及肺部病理学检测和气道反应性测定等方法检测NF κB顺式诱饵ODN对哮喘大鼠肺组织白细胞介素 5 (IL 5 )、诱导型一氧化氮合酶 (iNOS)mRNA和蛋白质表达及肺部炎症反应、气道反应性的影响。结果 ODN转染组大鼠肺部均有明显的FITC着色 ,说明此种局部滴入ODN的方法是可行的。经NF κB顺式诱饵ODN转染的哮喘大鼠 (N ODN组 )IL 5、iNOSmRNA表达均较未治疗的哮喘大鼠 (A组 )明显降低 (N ODNvsA ,0 .17±0 .0 3vs 0 .2 4± 0 .0 5 ,0 .2 4± 0 .0 5vs 0 .77± 0 .10 ,P <0 .0 5 ) ;蛋白质表达亦较A组减少 (40 .0± 10 .8vs80 .0± 2 5 .8,0 .19± 0 .0 4vs 0 .37± 0 .0 6 ,P <0 .0 5 ) ;且其肺部嗜酸粒细胞数量显著减少 (10 .0 0± 2 .94vs 16 .2 5± 4 .6 5 ,P <0 .0 5 ) ,气道反应性降低 (PC50 1.4± 0 .4vs0 .4± 0 .3,P <0 .0 5 )。而无序诱饵ODN转染的哮喘大鼠与未治疗的哮喘大鼠相比 ,上述指标差异无显著性 (P值均 >0 .0 5 )。结论 NF κB顺式诱饵元件整体给?

Objective To investigate the efficacy of cis element decoy against nuclear factor κB (NF-κB) binding site for treatment of asthma in vivo . Methods NF-κB decoy oligodeoxynucleotides (N-ODN) and scrambled decoy oligodeoxynucleotides (S-ODN) were introduced by administration into the lungs of asthmatic rats. The mRNA and protein levels of interleukin 5 (IL-5) and inducible nitric oxide synthase (iNOS) were measured by reverse transcription PCR, enzyme-linked immunosorbant assay and immunohistochemistry. The pathology of lung and airway responsiveness were also examined. Results ODN group showed a marked FITC-staining in the lung, demonstrating the efficacy of gene transfer into the lung by topical administration. N-ODN group showed a significantly lower expression of mRNA levels of IL-5 and iNOS (N-ODN group vs A group, 0.17±0.03 vs 0.24± 0.05 , 0.24±0.05 vs 0.77±0.10, P <0.05) and protein levels (40.0±10.8 vs 80.0±25.8,0.19±0.04 vs 0.37±0.06, P <0.05) than that in asthma group. At the same time, the N-ODN group showed significantly less eosinophil infiltration in the lung (10.00±2.94 vs 16.25±4.65, P <0.05) and significantly lower airway responsiveness as compared with asthma group (PC_ 50 1.4±0.4 vs 0.4±0.3, P <0.05). S-ODN had no significant differences with asthma group ( P >0.05). Conclusion Administration of NF-κB decoy ODN in lungs of rats with asthma inhibited expression of inflammatory mediators with amelioration of asthma. These findings suggest that NF-κB decoy ODN may function as a useful gene therapeutic approach in the treatment of asthma.