NF-κB介导哮喘过敏原刺激后支气管上皮细胞的凋亡

NF-κB mediates the apoptosis of the bronchial epithelial cells induced by asthma allergen

目的研究核转录因子κB(NF-κB)在哮喘过敏原刺激后支气管上皮细胞中的表达及对细胞凋亡的影响。方法以300U/L尘螨抗原提取物作为过敏原刺激支气管上皮细胞系16HBE细胞,应用RT-PCR和Western blot检测尘螨抗原提取物刺激对16HBE细胞NF-κB(p65)表达水平的影响;用NF-κB(p65)sh RNA沉默16HBE细胞内NF-κB表达后,应用流式细胞术和Western blot检测NF-κB(p65)在过敏原刺激诱导16HBE细胞凋亡中的作用。结果过敏原刺激后使16HBE细胞内NF-κB(p65)m RNA和蛋白水平明显上调,明显诱导16HBE细胞凋亡和活化型caspase-3水平上调,沉默NF-κB可使过敏原刺激诱导的16HBE细胞凋亡率降低和活化型caspase-3水平上调减少。结论 NF-κB可介导哮喘过敏原刺激后的支气管上皮细胞凋亡。

Objective To study the expression of NF-κB in bronchial epithelial cells after asthma allergen stimulation and its effect on cell apoptosis.Methods300U/L dust mite antigen extract was used as the allergen to stimulate bronchial epithelial cell line16HBE cells.The role of NF-κB(p65)in allergen induced16HBE cell apoptosis was explored through monitoring its expression level in response to allergen treatment as well as the apoptotic rate of16HBE cells after knocking down NF-κB using NF-κB(p65)shRNA.The NF-κB(p65)expression was determined by RT-PCR and Western blot.The apoptosis rate was measured using flow cytometry and Western blot.Results After allergen stimulation,NF-κB(p65)was significantly up-regulated in16HBE cells at both mRNA and protein levels.Increased apoptosis and caspase-3activation were observed in16HBE cells,which can be inhibited through silencing NF-κB.Conclusion NF-κB may contribute to the apoptosis of bronchial epithelial cells induced by asthma allergen.