目的:探讨中药复方制剂天龙咳喘灵提取物抗病毒损伤的可能机制。方法:运用短路电流技术、酶联免疫分析法和免疫荧光染色技术来研究天龙咳喘灵提取物对禽流感病毒膜蛋白血凝素(HA蛋白)所造成的气道损伤的恢复作用。结果:HA蛋白预处理能明显降低小鼠气道的基础分泌以及气道上皮细胞中cAMP的浓度,而天龙咳喘灵提取物的干预能在一定程度上使之得以恢复。另外,免疫荧光染色结果发现天龙咳喘灵提取物与HA蛋白共孵育能显著抑制NF-kB的入核。结论:天龙咳喘灵提取物能够活化cAMP依赖的囊性纤维化跨膜转导调节因子(CFTR通道),从而达到其抗病毒损伤的作用。
Objective:The aim of this study was to investigate the underlying antiviral mechanism of Tianlong Kechuanling(TLKCL)Extract in airway.Methods:Electrical measurements of mice tracheae and intracellular cAMP concentrations and the expression of NF-kB in cultured airway epithelial cells(16HBE)were detected respectively,which were treated with HA protein in the absence or presence of TLKCL.Results:Pretreatment with HA for 10min resulted in remarkable reduction at both the basal short-circuit current(Isc)of mice tracheae and cAMP level in the airway epithelial cells,which could be recovered in the presence of TLKCL.Moreover,addition of forskolin,an agonist of the cAMP-mediated Cl-channel(cystic fibrosis transmembrane conductance regulator,CFTR),increased Isc in HA-treated mice tracheae.Immunofluorescence staining revealed that the nucleic localization of NF-kB was reduced in 16HBE cells treated with HA and TLKCL compared with HA alone.Conclusion:The data suggests that TLKCL could recover HA protein-reduced secretion in airway via the activation of apical cAMP dependent CFTR Cl-channels.