芪仙汤提取物缓解苯并芘诱导的人气道上皮细胞黏蛋白5AC高表达的机制研究  

Mechanism of Qixian Decoction extract reversing Benzo(a)pyrene-induced Mucin 5AC expression in airway epithelial cells

作　　者：孙亦鹏 史兆雯 倪振华[1] 缪夏轶 林玉华 毕俊杰 王雄彪[1] SUN Yipeng;SHI Zhaowen;NI Zhenhua;MIAO Xiayi;LIN Yuhua;BI Junjie;WANG Xiongbiao(Department of Respiratory Medicine,Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200333,China)

机构地区：[1]上海中医药大学附属普陀医院呼吸科,上海200333

出　　处：《上海医学》2021年第4期251-256,共6页Shanghai Medical Journal

基　　金：上海市科学技术委员会科研计划项目(20ZR1450400,18140904000,17401970900),上海市普陀区卫生健康系统临床特色专科建设计划(2020tszk02),上海市普陀区自主创新科研项目/基本项目(ptkwws201714),上海市第六人民医院医疗集团课题(18-LY-01)。

摘　　要：目的探讨芪仙汤提取物对苯并芘(BaP)诱导的气道上皮细胞黏蛋白(MUC)5AC的抑制作用及其可能的机制。方法以人支气管黏液上皮细胞NCI-H292细胞株为研究对象,用BaP刺激细胞构建MUC5AC升高模型(模型组),分别用芪仙汤提取物240μg/mL(低浓度组)和480μg/mL(高浓度组)处理细胞株,空白组予二甲基亚砜(DMSO)处理。采用CCK-8法检测芪仙汤提取物处理后细胞活性的变化,实时荧光定量PCR检测MUC5AC和细胞色素P4501A1(CYP1A1)mRNA水平,免疫印迹法检测MUC5AC、CYP1A1、细胞外信号调节激酶(ERK)1/2、磷酸化ERK(p-ERK)1/2、相对分子质量为90000核糖体S6激酶(RSK)和磷酸化p90 RSK(p-p90 RSK)蛋白质表达水平,荧光探针检测细胞内活性氧(ROS)水平。结果浓度≤480μg/mL的芪仙汤提取物对NCI-H292细胞株活力无显著影响。与模型组相比,芪仙汤低、高浓度组MUC5AC蛋白质和MUC5AC mRNA水平均显著降低(P<0.05或0.01),ROS荧光强度值均显著降低(P<0.05或0.01),CYP1A1 mRNA水平均显著降低(P值均<0.01),p-ERK 1/2和p-p90 RSK蛋白质水平亦显著降低(P值均<0.01)。结论芪仙汤可下调BaP诱导的人气道上皮细胞MUC5AC基因和蛋白质高水平表达,其机制与抑制芳香烃受体活化介导的ROS和ERK通路激活相关。Objective To explore the effect of Qixian Decoction extract on the Benzo(a)pyrene(BaP)-induced mucin(MUC)5AC expression in airway epithelial cells and its possible mechanism.Methods Human bronchial mucus epithelial-like cells NCI-H292 was used as research object.BaP was applied as a stimulator to induce MUC5AC oversecretion.Then the cells were treated with Qixian Decoction extract 240μg/mL(low concentration)and 480μg/mL(high concentration).Dimethyl sulfoxide(DMSO)was given in blank control group.Cell counting kit 8(CCK-8)method was used to detect the proliferation of cells.Real time-polymerase chain reaction(PCR)was used to detect MUC5AC and cytochrome P450(CYP)1A1 mRNA levels.Western blot was used to detect MUC5AC,CYP1A1,extracellular signal-regulated kinase(ERK)1/2,phosphorylation ERK(p-ERK)1/2,RSK and p-p90 RSK protein expression.Fluorescence probes were used to detect intracellular levels of reactive oxygen species(ROS).Results Qixian Decoction extract(≤480μg/mL)had no obvious toxic effect on the viability of NCI-H292 cells.Compared with those in the model group,the protein and mRNA levels of MUC5AC in the low-and high-concentration Qixian Decoction group were significantly decreased(P<0.05 or 0.01);ROS level,the mRNA levels of CYP1A1,and the protein expressions of p-ERK 1/2 and p-p90 RSK in the low-and high-concentration Qixian Decoction group were also significantly decreased(P<0.05 or 0.01).Conclusion Qixian Decoction extract can inhibit BaP-induced MUC5AC protein and mRNA levels in airway epithelial cells.And the mechanism may be associated with the inhibition of ROS and ERK pathway activation mediated by AhR activation.

关 键 词：芪仙汤 黏蛋白质类 黏蛋白5AC 活性氧 苯并芘 芳香烃受体 

分 类 号：R285.5[医药卫生—中药学]