摘要
目的 探讨苏黄止咳胶囊中五味子醇甲对感冒后咳嗽(PIC)的改善作用.方法 气管滴注脂多糖(LPS)并联合香烟烟雾刺激建立体内PIC小鼠模型,小鼠随机分为对照组、模型组、苏黄止咳胶囊组(14 g/kg)、孟鲁司特钠组(阳性对照,3 mg/kg)和五味子醇甲低、高剂量组(10、30 mg/kg);LPS刺激人支气管上皮细胞(BEAS-2B)建立体外PIC模型,细胞分为对照组、模型组、苏黄止咳胶囊组(10 μg/mL)和五味子醇甲低、高浓度组(3、10 μmol/L).HE和Masson染色检测肺及支气管组织病理变化,ELISA法检测肺组织IL-1β、IL-6、TNF-α、ROS、MDA、SOD、GSH水平,RT-qPCR法检测BEAS-2B细胞IL-1β、IL-6、TNF-α mRNA表达,Western blot法检测小鼠肺组织和BEAS-2B 细胞中 p-PI3K、p-Akt、NOX4、SIRT1、p-ERK、Fibronectin、E-cadherin、Vimentin、α-SMA 蛋白表达.结果 与模型组比较,五味子醇甲和苏黄止咳胶囊均可延长PIC小鼠咳嗽潜伏期(P＜0.01),减少咳嗽次数(P＜0.01),减轻肺组织炎性细胞浸润和胶原沉积,降低肺组织 IL-1β、IL-6、TNF-α、ROS、MDA 水平和 Fibronectin、Vimentin、α-SMA、p-ERK、p-PI3K、p-Akt、NOX4 蛋白表达(P＜0.05,P＜0.01),升高肺组织SOD、GSH水平和E-cadherin、SIRT1 蛋白表达(P＜0.05,P＜0.01);降低BEAS-2B细胞ROS水平、IL-1β、IL-6、TNF-α mRNA表达和p-ERK、p-PI3K、p-Akt、NOX4 蛋白表达(P＜0.05,P＜0.01),升高BEAS-2B细胞SIRT1 蛋白表达(P＜0.01).结论 五味子醇甲可通过调节SIRT1/ERK信号通路抑制炎症,通过调节PI3K/Akt/NOX4 信号通路抑制氧化应激,发挥对PIC的止咳作用,是苏黄止咳胶囊的主要止咳功效成分之一.
Abstract
AIM To investigate the ameliorative effects of Schisandrol A(Sol A)in Suhuang antitussive capsule on post-infectious cough(PIC).METHODS The in vivo mouse PIC model was established by intratracheal instillation of lipopolysaccharide(LPS)combined with cigarette smoke exposure.The mice were randomly divided into the control group,the model group,the Suhuang antitussive capsule group(14 g/kg),the montelukast sodium positive control group(3 mg/kg),and low and high dose Sol A groups(10,30 mg/kg).The in vitro PIC model was established by stimulating human bronchial epithelial cells(BEAS-2B)with LPS.The cells were divided into the control group,the model group,the Suhuang antitussive capsule group(10 μg/mL)and low and high dose Sol A groups(3,10 μmol/L).HE and Masson staining were used to detect the pathological changes of the lung and bronchial tissues.ELISA was used to detect the levels of IL-1β,IL-6,TNF-α,ROS,MDA,SOD and GSH in the lung tissues.RT-qPCR was used to detect the IL-1β,IL-6 and TNF-α mRNA expressions in BEAS-2B cells.And Western blot was applied to detect the protein expressions of p-PI3K,p-Akt,NOX4,SIRT1,p-ERK,Fibronectin,E-cadherin,Vimentin and α-SMA in mouse lung tissue and BEAS-2B cells.RESULTS Compared with the model group,the groups intervened with Sol A or Suhuang antitussive capsule displayed prolonged cough latency(P＜0.01);reduced cough frequency(P＜0.01);relieved pulmonary inflammatory cell infiltration and collagen deposition in PIC mice;decreased pulmonary levels of IL-1β,IL-6,TNF-α,ROS,MDA and protein expressions of Fibronectin,Vimentin,α-SMA,p-ERK,p-PI3K,p-Akt,and NOX4(P＜0.05,P＜0.01);increased pulmonary levels of SOD and GSH and protein expressions of E-cadherin and SIRT1(P＜0.05,P＜0.01);decreased ROS level,IL-1β,IL-6,TNF-α mRNA expressions and p-ERK,p-PI3K,p-Akt,NOX4 protein expressions in vitro(P＜0.05,P＜0.01);and increased SIRT1 protein expression in vitro as well(P＜0.01).CONCLUSION Being the main antitussive component of Suhuang antitussive capsule upon the PIC model,Sol A inhibits the inflammation via SIRT1/ERK signaling pathway and relieve the oxidative stress via PI3K/Akt/NOX4 signaling pathway.
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