摘要
目的 探究噻托溴铵通过 MAPK/NF-κB 信号通路对哮喘的影响及其潜在的机制.方法 用 10 ng·mL-1 血小板衍生生长因子(platelet-derived growth factor,PDGF)-BB和不同浓度(0、5、10、20、50 μmol·L-1)噻托溴铵处理气道平滑肌细胞(airway smooth muscle cells,ASMCs),并通过 CCK-8 实验、克隆形成实验、划痕愈合实验和Transwell法分析 ASMCs细胞增殖、迁移和侵袭;Western blot 分析 MMP2、MMP9、calponin和α-SMA 的蛋白水平;Western blot检测 MAPK/NF-κB信号通路的水平.结果 噻托溴铵抑制 PDGF-BB诱导的 ASMCs 的增殖、迁移和侵袭(P＜0.05).噻托溴铵调节 ASMCs 的表型转变(P＜0.05).噻托溴铵可以抑制 PDGF-BB 诱导的MAPK/NF-κB信号通路激活,降低 p-38、p-JNK、p-ERK、p-p65、p-NF-κB的表达(P＜0.05).结论 噻托溴铵可以通过阻断 MAPK/NF-κB信号转导抑制 ASMCs的增殖和迁移.推测噻托溴铵可以作为一种治疗哮喘的有前途的药物.
Abstract
Objective To investigate the effect of tiotropium bromide on asthma through the MAPK/NF-κB signaling pathway and its potential mechanism.Methods Airway smooth muscle cells(ASMCs)were treated with 10 ng·mL-1 platelet-derived growth factor-BB(PDGF-BB)and different concentrations oftiotropium bromide(0,5,10,20 and 50 μmol·L-1).The prolifera-tion,colony formation ability,migration and invasion of ASMCs were analyzed by CCK-8,colony-formation,scratch wound healing and Transwell assays,respectively.The expression levels of MMP2,MMP9,calponin,α-SMAand MAPK/NF-κB signaling pathway proteins were analyzed by Western blot.Results Tiotropium bromide inhibited PDGF-BB-induced proliferation,migration and invasion and modulated phenotypic transformation in ASMCs(P＜0.05).Furthermore,tiotro-pium bromide inhibited DGF-BB-caused activation of MAPK/NF-κB signaling pathway,and re-duced the expression of p-38,p-JNK,p-ERK,p-p65 and p-NF-κB(P＜0.05).Conclusion Tiotro-pium bromide can inhibit the proliferation and migration of ASMCs by blocking MAPK/NF-κB signal transduction.It is speculated that tiotropium bromide can be used as a promising drug for the treatment of asthma.
基金项目
陕西省医学科学研究课题计划(2019JM3334)
维普
万方数据