升麻素通过NF-κB通路对支气管上皮细胞过敏性炎症反应的抑制作用

Inhibitory effect of cimifugin on allergic inflammation in bronchial epithelial cells via NF-κB pathway

目的:探究升麻素通过NF-κB信号通路对屋尘螨(HDM)诱导的人支气管上皮细胞炎症损伤及上皮屏障功能的调控作用。方法:将人支气管上皮细胞BEAS-2B分为空白对照组(Control)、HDM组、HDM+升麻素(0.01μmol/L)组、HDM+升麻素(0.1μmol/L)组、HDM+升麻素(1μmol/L)组、阳性药物(HDM+地塞米松)组。MTT和TUNEL法分别检测细胞活力和凋亡;ELISA检测细胞炎症因子水平;跨上皮电阻(TEER)和FITC标记葡聚糖4 kD(FD-4)评估细胞单层膜的渗透性;免疫荧光检测细胞NF-κB核易位;Western blot分析凋亡、炎症、紧密连接及NF-κB信号通路相关蛋白表达水平。结果:HDM可诱导BEAS-2B细胞活力损伤,诱导细胞凋亡、炎症反应和上皮屏障损害,并激活NF-κB信号通路(均P<0.001)。升麻素处理可浓度依赖性地抑制暴露于HDM下BEAS-2B细胞的活力损伤(P<0.05)、细胞凋亡(P<0.01)、炎症反应(P<0.05)、上皮屏障损伤(P<0.05),并使NF-κB信号通路失活(P<0.05)。结论:升麻素可显著抑制HDM诱导的支气管上皮细胞炎症损伤和上皮屏障损伤,这一结果可为过敏性哮喘的防治提供新的策略。

Objective:To figure out the regulatory role of cimifugin in the inflammatory injury and epithelial barrier function in house dust mite(HDM)-induced human bronchial epithelial cells via NF-κB signaling.Methods:Human bronchial epithelial BEAS-2B cells were divided into blank control(Control)group,HDM group,HDM+cimifugin(0.01μmol/L)group,HDM+cimifugin(0.1μmol/L)group,HDM+cimifugin(1μmol/L)group and positive drug(HDM+Dex)group.Cell viability and apoptosis were respectively estimated by MTT and TUNEL assays.Levels of inflammatory factors in cells were examined by ELISA.Transepithelial electrical resistance(TEER)and FITC-dextran 40 kD(FD-40)flux assessed the permeability of cell monolayers.Nuclear translocation of NF-κB in cells was detected by immunofluorescence assay.Western blot was used to analysis expressions of apoptosis,inflammatory,tight junction and NF-κB signaling-associated proteins.Results:HDM induced viability injury,apoptosis,inflammatory response,epithelial barrier damage and activated NF-κB signaling(all P<0.001)in BEAS-2B cells.Cimifugin treatment dose-dependently inhibited the viability injury(P<0.05),apoptosis(P<0.01),inflammatory response(P<0.05),epithelial barrier damage(P<0.05)and inactivated NF-κB signaling(P<0.05)in BEAS-2B cells exposed to HDM.Conclusion:Cimifugin significantly inhibits HDM-elicited inflammatory injury and epithelial barrier damage in bronchial epithelial cells.This finding may provide novel strategies for the prevention and treatment of allergic asthma.