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吴茱萸水提物体外肝毒性的谱-毒关系研究
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| 篇名： | 吴茱萸水提物体外肝毒性的谱-毒关系研究 |
| TITLE： | Study on spectrum-toxicity relationship of in vitro hepatotoxicity of aqueous extract from Euodia rutaecarpa |
| 摘要： | 目的 研究吴茱萸水提物体外肝毒性的谱-毒关系。方法制备16批不同产地吴茱萸水提物。采用超高效液相色谱（UPLC）法以及《中药指纹图谱相似度评价系统（2012A版）》建立吴茱萸水提物的指纹图谱，并进行共有峰指认及相似度评价；以正常人肝细胞L02为对象，考察16批吴茱萸水提物对该细胞的抑制作用；采用灰色关联度法和偏最小二乘回归分析法分析吴茱萸水提物UPLC指纹图谱的共有峰与L02细胞肝毒性的谱-毒关系，并对与吴茱萸体外肝毒性相关性最大的色谱峰的对应成分进行分离、制备及鉴定。结果16批吴茱萸水提物共有27个共有峰，相似度为0.375～0.995；共指认出9个成分，分别为新绿原酸（峰5）、绿原酸（峰9）、隐绿原酸（峰10）、咖啡酸（峰12）、芦丁（峰16）、金丝桃苷（峰17）、去氢吴茱萸碱（峰19）、吴茱萸碱（峰24）、吴茱萸次碱（峰25）。16批吴茱萸水提物对L02细胞的生长均具有显著的抑制作用（P＜0.05或P＜0.01），抑制率在6.68％～67.95％之间。经灰色关联度分析发现，关联度大于0.8的共有峰有18个，由大到小依次为峰8＞峰3＞峰23＞峰7＞峰4＞峰9＞峰12＞峰2＞峰19＞峰6＞峰15＞峰5＞峰1＞峰17＞峰21＞峰26＞峰20＞峰14；经偏最小二乘回归分析发现，回归系数为正且变量重要性投影值大于1的共有峰有14个，按回归系数大小排序依次为峰8＞峰3＞峰23＞峰2＞峰7＞峰4＞峰12＞峰9＞峰19＞峰5＞峰17＞峰26＞峰10＞峰15。峰8与吴茱萸体外肝毒性相关性最大，进一步鉴定该峰对应的化学成分为6-O-反式咖啡酰葡萄糖酸。结论吴茱萸水提物的体外肝毒性作用是其多组分共同作用的结果，其中6-O-反式咖啡酰葡萄糖酸与其体外肝毒性相关性最大。 |
| ABSTRACT： | OBJECTIVE To study the spectru m-toxicity relationship of in vitro hepatotoxicity of aqueous extract from Euodia rutaecarpa. METHODS The aqueous extract from 16 batches of E. rutaecarpa from different habitats were prepared. The fingerprints of aqueous extract from E. rutaecarpa were established by ultra high performance liquid chromatography （UPLC） method and Similarity Evaluation System of TCM Fingerprint （2012A edition ），and common peaks were identified and the similarity was evaluated. Using normal human hepatocytes L 02 as subject ，inhibitory effect of aqueous extract from 16 batches of E. rutaecarpa to them were investigated. The spectrum-toxicity relationship of UPLC fingerprint of aqueous extract from E. rutaecarpa with the hepatotoxicity of hepatocytes L 02 was analyzed by grey relational analysis （GRA）and partial least squares regression analysis （PLSR）. The corresponding compound of the chromatographic peak with the greatest correlation with the in vitro hepatotoxicity of E. rutaecarpa were isolated ，prepared and identified. RESULTS There were 27 common peaks in UPLC fingerprints of aqueous extract from 16 batches of E. rutaecarpa ，with similarity of 0.375-0.995. Totally 9 peaks were confirmed ，i.e. neochlorogenic acid （peak 5），chlorogenic acid （peak 9），cryptochlorogenic acid （peak 10），caffeic acid （peak 12），rutin （peak 16），hyperin（peak 17），dehydroevotarine（peak 19），evotarine（peak 24），rutecarpine（peak 25）. The aqueous extract from 16 batches of E. rutaecarpa showed significant inhibitory effect on the growth of L 02 cells（P＜0.05 or P＜0.01），and the inhibitory rate ranged from 6.68％ to 67.95％. GRA showed that there were 18 common peaks with correlation degree greater than 0.8，which were peak 8＞peak 3＞peak 23＞peak 7＞peak 4＞peak 9＞peak 12＞peak 2＞peak 19＞peak 6＞ 4928381。E-mail：799247687@qq.com peak 15＞peak 5＞peak 1＞peak 17＞peak 21＞peak 26＞ peak 20＞peak 14 in descending order of correlation degree. PLSR showed that there were 14 peaks with regression coefficient＞0 and variable importance projection value ＞1，and the order of regression coefficient was peak 8＞peak 3＞peak 23＞ peak 2＞peak 7＞peak 4＞peak 12＞peak 9＞peak 19＞peak 5＞peak 17＞peak 26＞peak 10＞peak 15. Peak 8 had the greatest correlation with in vitro hepatotoxicity，and the corresponding compound of this peak was identified as 6-O-trans caffeoyl gluconic acid. CONCLUSIONS The in vitro hepatotoxicity of aqueous extract from E. rutaecarpa is the result of multiple component interaction，among which 6-O-trans caffeoyl gluconic acid shows closest relation with in vitro hepatotoxicity. |
| 期刊： | 2022年第33卷第01期 |
| 作者： | 刘舒凌，王剑，刘雯，黄凤玉，蒋东明，林晓彤，蒙已钦，李耀华 |
| AUTHORS： | LIU Shuling，WANG Jian，LIU Wen，HUANG Fengyu，JIANG Dongming，LIN Xiaotong，MENG Yiqin，LI Yaohua |
| 关键字： | 吴茱萸；指纹图谱；肝毒性；L02细胞；谱-毒关系；灰色关联度分析法；偏最小二乘回归分析法 |
| KEYWORDS： | Euodia rutaecarpa ；fingerprint；hepatotoxicity；L02 cell；spectrum-toxicity relationship ；grey relational analyses ； |
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