华细辛和北细辛HPLC特征图谱识别与抗炎靶点及抗炎成分分析

建立华细辛和北细辛HPLC特征图谱并结合聚类分析研究2种来源细辛的识别方法;应用网络药理学方法预测细辛潜在抗炎靶点并寻找潜在抗炎成分。对89批细辛药材(12批华细辛和77批北细辛)的数据进行分析确定了11个特征峰,用对照品、紫外光谱和LC-MS指认了11个特征成分。特征峰面积聚类分析显示华细辛和北细辛被分为2类,且利用特征峰面积比值可实现两者区分,当特征峰9(细辛素)/参照峰S(卡枯醇)峰面积比值大于5时为华细辛,小于2时为北细辛。对119种细辛成分进行网络药理学分析的结果表明细辛抗炎作用可能与COX-2,COX-1,iNOS,MAPK14,LAT4H,NR3C1,PPARG和TNF等8个靶点相关,其中COX-2最为关键,与5种特征成分细辛脂素、芝麻脂素、细辛素、甲基丁香酚和黄樟醚均存在相互作用。此外,细辛脂素、芝麻脂素与iNOS,MAPK14也存在相互作用关系,黄樟醚和细辛素可作用于iNOS,COX-1,LAT4H,甲基丁香酚可作用于COX-1,LAT4H。细辛脂素与芝麻脂素均可作用在COX-2,iNOS和MAPK143个靶点上,提示它们是细辛发挥抗炎作用的活性成分;COX-2分子对接结果和COX-2活性实验结果验证了细辛脂素、芝麻脂素可抑制COX-2活性,为细辛抗炎作用活性成分。基于HPLC特征图谱的华细辛和北细辛识别方法简便易行;预测到的细辛抗炎靶点和抗炎成分为完善细辛质量评价体系奠定了基础。

Identification based on HPLC and anti-inflammatory targets as well as related constituents analysis of Asarum heterotropoides var.mandshuricum and A.sieboldii

The present work is to establish an HPLC characteristic chromatograms of Asarum heterotropoides var.mandshuricum(AH)and A.sieboldii(AS),combined with cluster analysis for the identification of the two species,and predict their potential anti-inflammatory related targets by network pharmacological method.Eighty-nine samples(12 batches of AS and 77 batches of AH)were analyzed,and 11 characteristic peaks were identified by reference substances,UV spectrum and LC-MS.Cluster analysis showed that AS and AH were divided into two groups,and the ratio of characteristic peak areas can be used to distinguish them.When the ratio of characteristic peak sarisan to kakuol was greater than 5,it was AS,and when the ratio was less than 2,it was AH.The network pharmacological analysis of 119 constituents of Asari Radix et Rhizoma suggested that the anti-inflammatory effect of Asari Radix et Rhizoma might be related to COX-2,COX-1,iNOS,MAPK14,NR3 C1,PPARG and TNF.Among them,COX-2 is a relatively key target,which interacted with the characteristic constituents,asarinin,sesamin,safrole,methyleugenol and sarisan.The characteristic constituents asarinin and sesamin also interacted with the iNOS and MAPK14.Safrole and sarisan can also interact with iNOS,COX-1 and LAT4 H.Methyleugenol also showed interaction with COX-1 and LAT4 H.Since asarinin and sesamin interacted with three targets,COX-2,iNOS and MAPK14,it implied that they were the main active constituents for the anti-inflammatory activity of Asari Radix et Rhizoma.The COX-2 inhibitory activities of asarinin and sesamin were further studied by molecular docking and bioassay.The HPLC method established was simple,feasible and reliable,with predicted anti-inflammatory targets and anti-inflammatory constituents,which could provide a reference for improving the quality evaluation system of Asari Radix et Rhizoma.