基于谱效关系的中药虎杖抗炎活性成分探讨  

Anti-inflammatory Active Ingredient of Chinese Herb Polygoni Cuspidati Rhizoma et Radix Based on Spectrum-effect Relationship

作　　者：李姣姣 于猛[2] 高源[2] 张涛[2] 贾红梅[2] 张宏武[2] 马丽焱[2] 邹忠梅[2] LI Jiao-jiao;YU Meng;GAO Yuan;ZHANG Tao;JIA Hong-mei;ZHANG Hong-wu;MA Li-yan;ZOU Zhong-mei(Tianjin University of Traditional Chinese Medicine, Tianjin 300193,China;The Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China)

机构地区：[1]天津中医药大学,天津300193 [2]中国医学科学院北京协和医学院药用植物研究所,北京100193

出　　处：《中国实验方剂学杂志》2019年第12期208-213,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家"重大新药创制"科技重大专项(2018ZX09711001-008);中国医学科学院医学与健康科技创新工程项目(2016-I2M-3-015)

摘　　要：目的:基于谱效关系研究虎杖中潜在的抗炎活性成分。方法:应用脂多糖(lipopolysaccharide,LPS)诱导小鼠腹腔巨噬细胞(RAW264. 7细胞)建立炎症模型,对虎杖95%乙醇提取物及其大孔树脂洗脱部位(30%,60%,95%乙醇洗脱物)的抗炎活性进行筛选;结合超高效液相色谱-飞行时间质谱(ultra performance liquid chromatography coupled with quadrupole timeof-flight mass spectrometry,UPLC-Q-TOF/MS)技术建立特征指纹图谱;利用偏最小二乘法(partial least squares,PLS)将各部位特征成分谱峰面积与一氧化氮(NO)抑制率进行谱效关系研究,根据变异权重系数(variable important in projection,VIP)来辨识其抗炎活性成分。结果:虎杖醇提物大孔树脂60%乙醇洗脱部位对LPS诱导的细胞炎症模型中一氧化氮生成的抑制能力最强,且表现出一定的剂量依赖关系。谱效关系研究发现,虎杖醇提物及其大孔树脂洗脱部位中3个成分包括大黄素-8-O-β-D-葡萄糖苷(E-8-G),大黄素-1-O-β-D-葡萄糖苷和大黄素-8-O-(6'-O-丙二酰基)-葡萄糖苷具有潜在的抗炎活性,并从细胞层面和分子反向寻靶研究对E-8-G的抗炎活性进行了验证。结论:本研究基于谱效关系的研究方法,从虎杖中筛选到3个潜在的抗炎活性成分,该方法有助于快速准确地发现中药活性成分(群),为研究中药药效物质基础提供新的研究思路和方法。Objective: To study the potential anti-inflammatory active ingredient of Chinese herbal Polygoni Cuspidati Rhizoma et Radix. Method: The inflammatory model of peritoneal macrophages( RAW264. 7 cells) induced by lipopolysaccharide( LPS) in mice was used to screen out the anti-inflammatory activity of 95%ethanol extract of Polygoni Cuspidati Rhizoma et Radix and its macroporous resin elution site( 30%,60%,95%ethanol eluting site). The characteristic fingerprints were established by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry( UPLC-Q-TOF/MS) technology,and then partial least squares method( partial least squares,PLS) was used to study the spectrum-effect relationship between the peak area of the characteristic components and the inhibition rate of nitric oxide( NO),and potential anti-inflammatory active ingredients were identified according to variable important in projection( VIP). Result: The ethanol extract macroporous resin 60% ethanol elution site of Polygoni Cuspidati Rhizoma et Radix had the strongest inhibition ability of nitric oxide formation, with a certain dose-dependent relationship. The study of spectrum-effect relationship showed that 3 components had potential anti-inflammatory activity,namely Emodin-8-O-β-D-glucoside( E-8-G),Emodin-1-O-β-D-giucoside,and Emodin-8-O-( 6’-O-malonyl)-glucoside. In addition,the antiinflammatory activity of E-8-G was further validated at the cell level through molecular docking analysis.Conclusion: Three potential anti-inflammatory active ingredients were found base on the spectrum-effect relationship. This study strategy is helpful to find the active ingredients( group) of traditional Chinese medicine,and provides new research ideas and methods for studying the material basis of Chinese herbal medicine.

关 键 词：虎杖 抗炎活性 谱效关系 超高效液相色谱-飞行时间质谱 大黄素-8-O-β-D-葡萄糖苷(E-8-G) 

分 类 号：R2-0[医药卫生—中医学] R22