目的 研究仲景名方白头翁汤的化学成分,建立HPLC指纹图谱,并结合网络药理学对其功效关联物质进行机制预测分析,为白头翁汤质量控制提供参考依据。方法 采用超高效液相色谱-四极杆飞行时间质谱联用技术(UPLC-Q-TOF-MS/MS)对白头翁汤的化学成分进行定性分析,建立HPLC指纹图谱,进行相似度评价,匹配共有峰并进行色谱峰归属。基于质谱鉴定和指纹图谱筛选白头翁汤的有效成分,通过网络药理学建立了"组分-靶点-通路"网络,进一步佐证白头翁汤有效成分选择的合理性。结果 通过数据库匹配、元素组成和碎片结构分析,共鉴定出21种化合物;在建立15批白头翁汤样品的指纹图谱中,标定16个共有峰,通过对照品指认出秦皮甲素(峰2)、秦皮乙素(峰5)、秦皮素(峰7)、白头翁皂苷B4(峰15)、白头翁皂苷B5(峰16)、黄柏碱(峰3)、黄连碱(峰11)、小檗碱(峰13)、巴马汀(峰14)9个色谱峰,各批次样品相似度均>0.90,且16个共有峰均能归属到白头翁、黄连、黄柏、秦皮4味药材;通过网络药理学对所指认的9个药效成分进行机制预测,富集的通路中含癌症通路、磷脂酰肌醇3-激酶/蛋白激酶B(phosphatidylinositol-3-kinase/protein kinase B,PI3K/Akt)信号通路以及缺氧信号转导-1(hypoxia signaling-1,HIF-1)信号通路等,这些通路可能与白头翁汤治疗疾病的作用相关。结论 UPLC-Q-TOF-MS/MS法能快速鉴定白头翁汤的成分,建立的白头翁汤指纹图谱分析方法简便可行,结合网络药理学发现白头翁汤9个成分与白头翁汤功效属性密切相关,为白头翁汤的质量控制提供参考,同时也为白头翁汤药效关联物质基础的研究及作用机制的探索奠定基础。

Objective To study the chemical components of a famous prescription of Zhongjing named Baitouweng Decoction (白头翁汤), establish HPLC fingerprint and analyze the mechanism of its efficacy-related substances using network pharmacology, so as to provide a reference for the quality control of Baitouweng Decoction. Methods Qualitative analysis of the chemical components of Baitouweng Decoction was carried out by UPLC-Q-TOF-MS/MS, the HPLC method was used to establish the fingerprint analysis method of Baitouweng Decoction and the similarity was evaluated. The common peaks were matched, at the same time, the attribution analysis of each common peak was performed. The active ingredients of Baitouweng Decoction were screened based on Mass spectrometry identification and fingerprints, and the "component-target-pathway" network was established through network pharmacology, which further supported the rationality of functional components selection for Baitouweng Decoction. Results Through database matching, element composition and fragment structure analysis, a total of 21 compounds were identified. The Baitouweng Decoction fingerprint analysis method was established, 15 batches of Baitouweng Decoction samples were measured, 16 common peaks were calibrated, and nine peaks of esculin (peak 2), aesculetin (peak 5), fraxetin (peak 7), anemoside B4 (peak 15), anemoside B5 (peak 16), phellodendrine (peak 3), coptisine (peak 11), berberine (peak 13) and palmatine (peak 14) were identified by comparing with the reference substance. The similarity of each batch of samples was greater than 0.90. And the 16 common peaks can be assigned to the four flavors of Baitouweng (Pulsatillae Radix), Huanglian (Coptidis Rhizoma), Huangbo (Phellodendri Chinensis Cortex), and Qinpi (Fraxini Cortex). The mechanism of the identified nine medicinal components was predicted. The enriched pathways included cancer pathways, PI3K-Akt signaling pathways, and HIF-1 signaling pathways, which may be related to the effect of Baitouweng Decoction in treating diseases. Conclusion The UPLC-Q-TOF-MS/MS method can quickly identify the chemical components of Baitouweng Decoction, and the fingerprint analysis method established in this experiment is simple and feasible. Combined with network pharmacology, it is found that the nine components of Baitouweng Decoction are closely related to the efficacy and attributes of Baitouweng Decoction, which provides a reference for the overall control of the quality and provides a material basis for the efficacy of Baitouweng Decoction. The research and the exploration of the mechanism of action also lay the foundation.

R283.6

国家自然科学基金资助项目（81860711）；国家自然科学基金资助项目（81860270）；广西自然科学基金资助项目（2018GXNSFBA281034）；广西科技基地和人才专项（桂科AD19245124）