基于谱效关系的灯盏细辛体外抗血小板聚集活性成分研究  

Study on antiplatelet aggregation active components of Erigeron breviscapus in vitro based on spectrum-effect correlation analysis

作　　者：伍珊娜 刘振杰 章从恩[2] 何琴 王玄[3] 牛明[2] 王伽伯[2] 肖小河[2] 

机构地区：[1]江西中医药大学,江西南昌330004 [2]解放军第三0二医院全军中医药研究所,北京100039 [3]首都医科大学中医药学院,北京100069

出　　处：《中草药》2017年第24期5179-5185,共7页Chinese Traditional and Herbal Drugs

基　　金：全军医学科技“十二五”科研项目:中国军事本草的调研整理与推广应用(BWS11J049,负责人:肖小河);20种道地药材优良品种质量生物评价技术研究(201507002,负责人:肖小河);第三〇二医院院长创新基金:便携式野战中医药医疗箱研制(QNPY2015021,负责人:牛明)

摘　　要：目的通过生物活性检测结合化学指纹图谱分析,探索灯盏细辛抗血小板聚集的活性成分。方法采用UPLC-UV分析技术建立不同批次灯盏细辛药材化学指纹图谱,对不同批次灯盏细辛进行抗血小板聚集活性效价检测,基于谱效关系推测可能的活性物质,并对5个高相关性成分进行体外抗血小板聚集作用的验证,根据5种单体化合物在灯盏细辛中的含量差异,计算5个单体化合物的相对活性贡献度。结果通过化学指纹图谱与抗血小板聚集生物效价的谱效相关分析,筛选并鉴定出与生物活性相关系数大于0.5的5个色谱峰,分别鉴定为绿原酸、咖啡酸、野黄芩苷、异绿原酸A、异绿原酸C。进一步体外实验表明5个化合物在相同质量浓度下均有不同程度的抗血小板聚集作用(抑制率16.5%~85.5%),相对活性强度顺序:野黄芩苷>异绿原酸C>咖啡酸>异绿原酸A>绿原酸;而5种成分从相对活性贡献度来看,异绿原酸C与野黄芩苷的活性贡献度大于另外3种成分。结论建立了灯盏细辛体外抗血小板聚集生物效价的检测方法;且野黄芩苷和异绿原酸C是灯盏细辛体外抗血小板聚集的主要活性成分。Objective To explore the active components of antiplatelet aggregation of Erigeron breviscapus, chemical fingerprints and bioactivity detection were used to carry out spectrum-effect correlation analysis. Methods A fingerprinting method was established by ultra-high performance liquid chromatography with ultraviolet detection（UPLC-UV） and then used for fingerprinting of different batches of E. breviscapus. The antiplatelet aggregation biopotency of different batches of E. breviscapus was tested, and the possible active substances were deduced based on spectrum-effect correlation analysis. Furthermore, all five compounds were verified by antiplatelet aggregation in vitro, and the contribution value of relative activity of the five compounds was calculated according to the difference of the contents of five kinds of monomer compounds in E. breviscapus. Results Through the spectrum-effect correlation analysis of chemical fingerprints and antiplatelet aggregation biopotency of E. breviscapus, five chromatographic peaks with higher bioactivity correlation coefficient were screened and identified, including chlorogenic acid, caffeic acid, scutellarin, isochromic acid A, and ischlorogenic acid C. Further in vitro experiments showed that the five compounds had different levels of antiplatelet aggregation at same concentration（inhibition rate： 16.5%—85.5%）. The sequence of the relative activity contribution is scutellarin ischlorogenic acid C caffeic acid ischlorogenic acid A chlorogenic acid. In terms of activity contribution of five compounds, chlorogenic acid C and scutellarin was larger than other compounds. Conclusion A method for the determination of antiplatelet aggregation biopotency of E. breviscapus in vitro was established. Moreover, scutellarin and ischlorogenic acid C are the main active substances of E. breviscapus in the aspect of antiplatelet aggregation in vitro.

关 键 词：灯盏细辛 化学指纹图谱 抗血小板聚集 生物效价 谱效相关 绿原酸 咖啡酸 野黄芩苷 异绿原酸A 异绿原酸C 

分 类 号：R285.5[医药卫生—中药学]