基于指纹图谱和网络药理学对曼地亚红豆杉质量标志物的分析研究  

Analysis of quality markers of Taxus media based on fingerprint and network pharmacology

作　　者：李雅静 张振凌[1,2,3,4] 王胜超 朱建光[1,2] 王瑞生 李宝卿[5] 赵永琪 孙梦梅 LI Ya-jing;ZHANG;Zhen-lingg;WANG Sheng-chao;ZHU Jian-guang;WANG Rui-sheng;LI Bao-qing;ZHAO Yong-qi;SUN Meng-mei(College of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Research Center for Special Processing Technology of Chinese Medicine,Zhengzhou 450046,China;Co-construction Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases by Henan&Education Ministry of P.R,Zhengzhou 450046,China;Henan Provincial Key Laboratory of TCM Decoction Processing,Zhengzhou 450046,China;Henan Jing Chun-Tang Pharmaceutical Industry Company Limited,Xuchang 461000,China)

机构地区：[1]河南中医药大学药学院,郑州450046 [2]河南省中药特色炮制技术工程研究中心,郑州450046 [3]呼吸疾病中医药防治省部共建协同创新中心,郑州450046 [4]河南省中药饮片炮制中医药重点实验室,郑州450046 [5]河南省景春堂药业有限公司,许昌461000

出　　处：《药物分析杂志》2024年第1期35-50,共16页Chinese Journal of Pharmaceutical Analysis

基　　金：河南省药品监督管理局科技计划项目(2020DB224);河南中医药大学2021年度科技协同创新专项项目(2023XTCX041);河南中医药大学2022年度研究生科研创新项目(2022KYCX006);公益性行业专项—中药炮制技术传承基地建设(00104296)。

摘　　要：目的:基于指纹图谱和网络药理学方法,对曼地亚红豆杉质量标志物(Q-Marker)进行预测,建立基于Q-Marker的曼地亚红豆杉药材评价方法。方法:采用Waters SymmmetryShield^(TM)RP 18(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.1%三氟乙酸水为流动相,梯度洗脱,流速1 mL·min^(-1),柱温30℃,检测波长254 nm,进样体积10μL,建立8个年限24批曼地亚红豆杉药材指纹图谱并标定共有峰,采用层次聚类分析(hierarchical clustering analysis,HCA)进行分类评价,借助正交偏最小二乘法-判别分析(orthogonal partial least square-discriminant analysis,OPLS-DA)筛选不同年限曼地亚红豆杉的主要差异性标志物;结合网络药理学,通过相应数据库筛选差异性标志物的核心靶点和关键通路,构建“成分-靶点-通路”网络图,以此预测曼地亚红豆杉的Q-Marker;采用HCA及主成分分析(principal component analysis,PCA)进一步验证曼地亚红豆杉的Q-Marker,综合评价其质量。结果:建立了24批曼地亚红豆杉药材指纹图谱,标定了25个共有峰,指认出18个化合物,包括紫杉烷类、黄酮类、生物碱类、甾体类、酚类,相似度均>0.900;HCA的结果显示不同年限曼地亚红豆杉存在一定的差异性;经OPLS-DA筛选出11个差异性标志物,分别为紫杉醇、10-脱乙酰基巴卡亭Ⅲ(10-DAB)、巴卡亭Ⅲ、三尖杉宁碱、10-去乙酰紫杉醇(10-DAT)、阿魏酸、山柰素、芦丁、金松双黄酮、穗花杉双黄酮、对羟基苯甲醛;基于差异性标志物,运用网络药理学从有效性角度对其进行分析,初步预测紫杉醇、10-DAB、巴卡亭Ⅲ、三尖杉宁碱、10-DAT为曼地亚红豆杉的Q-Marker,主要通过作用于PIK3R1、AKT1、EGFR、HRAS、MAPK1等14个核心靶点,调控癌症通路、AGE-RAGE信号在糖尿病并发症中的作用等信号通路,发挥消肿散结、通经利尿的功效;HCA结果验证了预测的Q-Marker的合理性,PCA综合评价结果表明十年生春季采收的曼地亚红豆杉质�Objective:To predict potential quality markers(Q-Marker) of Taxus media Rehd. based on fingerprint and network pharmacology methods, and establish an evaluation method of Taxus media Rehd. based on Q-Marker, as so to provide basis for the establishment of quality standard and quality evaluation system of Taxus media Rehd.. Methods: Waters SymmmetryShield^(TM )RP18(250 mm×4.6 mm, 5 μm) chromatographic column was used for separation, and the wavelength was 254 nm. The mobile phase was acetonitrile-0.01%trifluoroacetic acid aqueous solution for gradient elution at a flow rate of 1.0 mL·min^(-1), and the column temperature was 30 ℃. The injection volume was 10 μL. The fingerprints of 24 batches of Taxus media Rehd. with 8 kinds of cultivation years were established and the common peaks were identified. Hierarchical clustering analysis(HCA) was used for classification, orthogonal partial least square-discriminant analysis(OPLS-DA) was used to screen out the main marker components that cause differences between different years. Combined with network pharmacology, the core targets and key pathways were constructed a “component-target-pathway” network map through corresponding databases. Q-Markers were further verified and quality of Taxus media Rehd. comprehensively evaluated by HCA and PCA. Results: A total of 25 common peaks were obtained in the HPLC fingerprint of Taxus media Rehd., 18 compounds were identified, and their similarities were all above 0.900, including taxanes, flavonoids, alkaloids, steroids and phenols. The results of HCA showed that there were some differences among samples with different cultivation years. Eleven different markers were screened by OPLS-DA, and they were paclitaxel, 10-deacetylbaccatin Ⅲ(10-DAB), baccatin Ⅲ, cephalomannine, deacetyltaxol(10-DAT), ferulic acid, kaempferide, rutin, amentoflavone-4',4",7-trimethyl ether, skyrin, 4-hydroxybenzaldehyde. Based on the difference markers, network pharmacology was applied to analyze them from the perspective of effectiveness. Pa

关 键 词：曼地亚红豆杉 指纹图谱 网络药理学 化学识别模式 紫杉醇 10-脱乙酰基巴卡亭Ⅲ 巴卡亭Ⅲ 三尖杉宁碱 10-去乙酰紫杉醇 质量标志物 质量评价 

分 类 号：R917[医药卫生—药物分析学]