噻托溴铵对乙酰胆碱诱导的支气管上皮细胞NF-κB活化和IL-8表达的影响

Effect of tiotropium on activity of NF-κB and the expression of IL-8 in acetylcholine-stimulated human bronchial epithelial cells

目的观察噻托溴铵对乙酰胆碱诱导的支气管上皮细胞(16HBE)分泌IL-8及核转录因子NF-NF-κB活化的影响。方法使用不同浓度Ach、噻托溴铵和乙酰胆碱+噻托溴铵等分别处理生长良好的支气管上皮细胞(16HBE),用ELISA法检测细胞及其上清液中IL-8的表达变化,用Western Blot法检测细胞核内NF-κB蛋白含量变化。结果与对照组相比,乙酰胆碱能刺激16HBE分泌IL-8且呈剂量依赖,并在10 uM浓度时达到刺激高峰(P<0.001)。这种刺激作用与乙酰胆碱诱导16HBE的NF-κB活化有关(P<0.05)。噻托溴铵预处理可抑制乙酰胆碱诱导的NF-κB活性(P<0.05)及IL-8分泌(P<0.05)。结论噻托溴铵通过阻断NF-κB的活化,下调乙酰胆碱诱导16HBE细胞分泌IL-8水平,可能是其治疗COPD的抗炎作用机制之一。

Objective To investigate the inhibitory effects of tiotropium on the activation of nuclear factor kappa B（NF-κB） and the expression of interleukin-8（IL-8） in human bronchial epithelial cell line（16HBE） induced by acetylcholine and to discuss the possible anti-inflammatory mechanisms of tiotropium on the treatment of chronic obstructive pulmonary disease（COPD）.Methods Human bronchial epithelial cell line（16HBE） were cultured in vitro,and treated with different concentration of acetylcholine（Ach） in the presence or absence of tiotropium for different time.The protein expression of IL-8 were determined by Elisa.The expression of NF-κB in the nuclei was determined by Western Blot.Results The expression of IL-8 were markedly up-regulated and the expression of NF-κB also significantly increased in 16HBE stimulated with Ach compared with the control group,and 10 uM concentrations to stimulate the peak（P0.05）.Tiotropium pretreatment inhibited acetylcholine-induced NF-κB activity（P0.05） and IL-8 secretion（P0.05）.Conclusions Tiotropium effectively down-regulated IL-8 expression in 16HBE by inhibiting the activation of NF-κB,which may be one of its anti-inflammatory actions.