半夏水煎液不同部位拮抗小鼠呼吸道炎症作用及其效应部位成分分析  

Effect of different parts of Pinelliae Rhizoma Decoction against airway inflammation and analysis of effective components

作　　者：薛凡 郁红礼[1,2,3] 刘睿[1,4,5] 吴皓[1,2,3] 张元斌 刘东方 张萍[6] 陈盛君[7] 李松[7] XUE Fan;YU Hong-li;LIU Rui;WU Hao;ZHANG Yuan-bin;LIU Dong-fang;ZHANG Ping;CHEN Sheng-jun;LI Song(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Key Laboratory of Chinese Medicine Processing,Nanjing 210023,China;Engineering Research Center of Normalization and Standardization of Processing of Chinese Materia Medica,Nanjing 210023,China;Research and Development Center of Marine Drugs of Jiangsu Province,Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,Nanjing University of Chinese Medicine,Nanjing 210023,China;National Institutes for Food and Drug Control,Beijing 100050,China;Jiangyin Tianjiang Pharmaceutical Co.,Ltd,Jiangyin 214400,China;Jiashan Hospital of Traditional Chinese Medicine,Jiaxing 314100,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]江苏省中药炮制重点实验室,江苏南京210023 [3]国家教育部中药炮制规范化及标准化工程研究中心,江苏南京210023 [4]南京中医药大学江苏省海洋药用生物资源研究与开发重点实验室,江苏南京210023 [5]南京中医药大学江苏省中药资源产业化过程协同创新中心,江苏南京210023 [6]中国食品药品检定研究院,北京100050 [7]江阴天江药业有限公司,江苏江阴214400 [8]嘉善县中医医院,浙江嘉兴314100

出　　处：《中国中药杂志》2021年第22期5912-5921,共10页China Journal of Chinese Materia Medica

基　　金：国家重点研发计划项目(2018YFC1707000);国家自然科学基金项目(81573605)

摘　　要：探究半夏水煎液治疗呼吸道炎症的物质基础及作用机制。采用香烟烟雾联合脂多糖(lipopolysaccharide,LPS)诱导的小鼠呼吸道炎症模型,以小鼠肺泡灌洗液(broncho-alveolar lavage fluid,BALF)中炎症因子IL-6、IL-8表达水平,肺组织p38、IκB蛋白及其磷酸化水平为指标;采用香烟提取物(cigarette smoke extraction,CSE)诱导的支气管上皮细胞(human bronchial epithelial cell,16-HBE)损伤模型,以胞内IL-6、IL-8 mRNA表达水平及炎症信号通路关键蛋白p38、IκB蛋白及其磷酸化水平为指标,评价半夏水煎液不同部位的抗炎作用。结果表明,半夏水煎液具有显著的拮抗小鼠呼吸道炎症的作用,能下调呼吸道炎症小鼠及CSE诱导16-HBE细胞炎症因子IL-6、IL-8表达水平,抑制p38、IκB蛋白磷酸化;半夏水煎液的二氯甲烷、乙酸乙酯部位具有显著的拮抗炎症作用,其他部位无明显作用。同时构建半夏成分结构数据库,采用HPLC-TOF-MS和Nano-LC-MS/MS对效应部位成分进行成分组成分析,结果显示2个效应部位的成分组成类型相似,故合并2个部位后以阳离子交换树脂分为生物碱部位与非生物碱部位。体外效应研究结果显示,半夏的非生物碱部位具有显著的抗炎效应,但生物碱部位无明显作用。质谱分析结果显示,半夏非生物碱部位含有172种多肽和7种有机酸。以BCA法测定半夏非生物碱部位中肽质量分数为63.5%,以电位滴定法测定有机酸质量分数为9.92%。半夏水煎液可通过抑制p38、IκB磷酸化,阻断MAPK、NF-κB信号通路活化发挥拮抗小鼠呼吸道炎症作用,其效应成分可能与其所含的肽类、有机酸类成分相关。This study investigated the material basis and mechanism of Pinelliae Rhizoma Decoction in the treatment of airway inflammation.The cigarette smoke combined with lipopolysaccharide(LPS)was used to induce an airway inflammation model in mice.The expression levels of IL-6 and IL-8 in the bronchoalveolar lavage fluid(BALF)and the phosphorylation levels of p38 and IκB in the lungs of mice were taken as indexes to screen the effective extracts by system solvent extraction from Pinelliae Rhizoma Decoction(dichloromethane extract,ethyl acetate extract,n-butanol extract,etc.).Meanwhile,the human bronchial epithelial(16-HBE)cell model of cigarette smoke extract(CSE)-induced injury was established,and the mRNA expression levels of IL-6 and IL-8 and the phosphorylation levels of p38 and IκB proteins were also taken as indexes to evaluate the anti-inflammatory effect of different extracts of Pinelliae Rhizoma Decoction.The results showed that Pinelliae Rhizoma Decoction significantly antagonized airway inflammation in mice by down-regulating the expression levels of IL-6 and IL-8 in mice with airway inflammation and 16-HBE cells with CSE-induced injury and inhibiting the phosphorylation levels of p38 and IκB.The dichloromethane and ethyl acetate extracts of Pinelliae Rhizoma Decoction showed significant anti-inflammatory effects,while such effects of other extracts were not prominent.Furthermore,the database of Pinelliae Rhizoma composition was constructed,and the components in effective extracts were analyzed by HPLC-TOF-MS and Nano-LC-MS/MS.As revealed by the results,the compositions of the two effective extracts were similar with 36 common components.They were combined and then divided into Pinelliae Rhizoma alkaloids(PTAs)and Pinelliae Rhizoma non-alkaloids(PTNAs)by 732 cation-exchange resin.Further in vitro investigation confirmed the significant anti-inflammatory effect of PTNAs,while such effect of PTAs was not manifest.The MS analysis showed 172 peptides and 7 organic acids in PTNAs.The peptide content in PTNAs wa

关 键 词：半夏 呼吸道炎症 p38 MAPK/NF-κB 非生物碱部位 

分 类 号：R285.5[医药卫生—中药学]