摘要
目的:本实验应用RNA干扰技术(RNA interference,RNAi)探讨沉默信号转导与转录激活因子1(signal transducerand activator of transcription 1,STAT1)对气道炎症的抑制作用。方法:利用前期实验构建的重组质粒(pGSTAT1)及阴性对照(pG-HK)转染活化STAT1蛋白表达的人支气道上皮细胞(16HBE),显微镜下观察细胞形态并确定转染效率,免疫印迹法检测STAT1及细胞间黏附分子1(ICAM1)蛋白表达,直线相关分析两种蛋白之间相关性。结果:IFN-γ刺激16HBE细胞30min可以引起STAT1表达增加,pGSTAT1重组质粒可以有效转染刺激后的16HBE细胞,并使STAT1及ICAM1蛋白表达明显降低,且两种蛋白的表达存在正相关。结论:pGSTAT1重组质粒明显下调STAT1及ICAM1蛋白的表达,从而可能有效抑制气道炎症,为利用RNA干扰技术治疗支气管哮喘提供了理论依据。
Objective：This study using RNA interference（RNAi） to explore the inhibition effect of the silent signal transducer and activator of transcription 1（STAT1） on airway inflammation.Methods： Human bronchial epithelial cells were cultured and divided into six groups,normal cells group,IFN-γ stimulated group,pG-HK group,pGSTAT1 24h group,pGSTAT1 48h group,and pGSTAT1 72h group.The total cellular protein of these groups were extracted respectively.STAT1 and intercellular adhesion molecule 1（ICAM1） protein expression were detected in the way of Western blot and the correlation between the two proteins were analyzed.Results： The pGSTAT1 can be effectively transfected 16HBE cells,STAT1 and ICAM1 protein expression were decreased sig-nificantly and there was positive correlation between STAT1 and ICAM1.Conclusion： This RNA interference technology may be used to inhibit airway inflammation,provide us a theoretical basis for RNAi treatment of bronchial asthma.
出处
《泸州医学院学报》
2010年第5期507-510,共4页
Journal of Luzhou Medical College
