目的 筛选五加皮治疗类风湿性关节炎（rheumatoid arthritis，RA）和骨质疏松（osteoporosis，OP）“异病同治”潜在质量标志物（quality marker，Q-Marker）。方法 采用超高效液相色谱-四极杆-飞行时间质谱（UHPLC-Q-TOF-MS）联用技术，鉴定五加皮提取物入血成分；通过网络药理学预测入血成分治疗RA和OP的共有核心靶点与关键通路，并采用分子对接技术，验证候选成分与核心靶点的结合能力；采用UHPLC技术，建立五加皮的指纹图谱及多成分含量测定方法，结合药材相似度评价及层次聚类分析，比较不同批次药材差异，基于Q-Marker五原则综合筛选潜在Q-Marker。结果 鉴定出五加皮提取物20种入血成分；经网络药理学分析，筛选出五加酸、长白楤木酸、异贝壳杉烯酸、无梗五加苷B、刺五加苷E、刺五加苷D、咖啡酸、异绿原酸B和异绿原酸C，可能通过调控白细胞介素-6（interleukin-6，IL-6）、肿瘤坏死因子（tumor necrosis factor，TNF）、前列腺素内过氧化物合酶2（prostaglandin-endoperoxide synthase 2，PTGS2）、半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，CASP3）、基质金属蛋白酶9（matrix metalloproteinase 9，MMP9）和过氧化物酶体增殖物激活受体γ（peroxisome proliferator-activated receptor gamma，PPARG）等20个核心靶点，参与环氧合酶-2（cyclooxygenase-2，COX-2）/表皮生长因子受体（epithelial growth factor receptor，EGFR）炎症轴和激素水平调控等相关通路；分子对接进一步证实，上述9个候选成分与6个关键核心靶点IL-6、TNF、PTGS2、CASP3、MMP9和PPARG具有良好结合活性。经UHPLC指纹图谱分析，指认五加皮中10个共有峰，其中3个可区别同属药材三加皮Acanthopanax trifoliatus的特征峰为刺五加苷B、刺五加苷D和五加酸，5个可区别五加叶（非药用部位）的特有峰为刺五加苷B、刺五加苷D、五加酸、长白楤木酸和8-O-4,8-O-4-去氢三阿魏酸。药材相似度评价结果显示，不同五加皮药材成分相近，多成分含量测定及层级聚类分析结果显示，10个成分在各批次药材中具有一定的差异。结合Q-Marker原则，最终筛选出五加酸、刺五加苷D、异贝壳杉烯酸、长白楤木酸、无梗五加苷B、咖啡酸、异绿原酸B、异绿原酸C和刺五加苷B为五加皮干预RA和OP“异病同治”的潜在Q-Marker。结论 五加酸、刺五加苷D、异贝壳杉烯酸、长白楤木酸、无梗五加苷B、咖啡酸、异绿原酸B、异绿原酸C和刺五加苷B为五加皮干预RA和OP“异病同治”的潜在Q-Marker。

Objective To screen the potential quality marker (Q-Marker) of Wujiapi (Acanthopanacis Cortex) for “same treatment for different diseases” in rheumatoid arthritis (RA) and osteoporosis (OP). Methods UHPLC-Q-TOF-MS was employed to identify blood-absorbed components of Acanthopanacis Cortex extract. Network pharmacology was used to predict common core targets and key pathways of blood-absorbed components in the treatment of RA and OP, and molecular docking was performed to verify the binding abilities between candidate compounds and core targets. UHPLC was applied to establish fingerprint and a multi-components quantitative analysis method, combined with similarity evaluation and hierarchical cluster analysis to compare differences between different batches of medicinal materials. Finally, potential Q-Markers were comprehensively screened based on the five principles of Q-Marker. Results A total of 20 blood-absorbed components were identified. Network pharmacology analysis predicted acanthoic acid, continentalic acid, kaurenoic acid, acanthoside B, eleutheroside E, eleutheroside D, caffeic acid, isochlorogenic acid B and isochlorogenic acid C as candidate components, which may participate in pathways related to the cyclooxygenase-2 (COX-2)/epithelial growth factor receptor (EGFR) inflammatory axis and hormone regulation by regulating 20 core targets, including interleukin-6 (IL-6), tumor necrosis factor (TNF), prostaglandin-endoperoxide synthase 2 (PTGS2), cystein-asparate protease-3 (CASP3), matrix metalloproteinase 9 (MMP9) and peroxisome proliferator-activated receptor gamma (PPARG). Molecular docking confirmed favorable binding activities between the nine candidate components and the six key targets, namely IL-6, TNF, PTGS2, CASP3, MMP9, and PPARG. Based on the UHPLC fingerprint analysis, 10 common peaks in Acanthopanacis Cortex were identified. Among these, three characteristic peaks (eleutheroside B, eleutheroside D, acanthoic acid) were effective in distinguishing Acanthopanacis Cortex from root bark of Acanthopanax trifoliatus of the same genus. In addition, five characteristic peaks, including eleutheroside B, eleutheroside D, acanthoic acid, continentalic acid and 8-O-4,8-O-4-dehydrotriferulic acid, were identified as potential chemical markers for differentiating Acanthopanax gracilistylus leaves (non-medicinal parts) from Acanthopanacis Cortex. The similarity evaluation results indicated that different batches of Acanthopanacis Cortex shared generally similar chemical compositions, whereas multicomponent content determination and hierarchical cluster analysis demonstrated certain differences in the contents of the 10 compounds among batches. Combined with the five principles of Q-Marker, acanthoic acid, eleutheroside D, kaurenoic acid, continentalic acid, acanthoside B, caffeic acid, isochlorogenic acid B, isochlorogenic acid C and eleutheroside B were ultimately screened as potential Q-Markers of Acanthopanacis Cortex for “same treatment for different diseases” in RA and OP. Conclusion Acanthoic acid, eleutheroside D, kaurenoic acid, continentalic acid, acanthoside B, caffeic acid, isochlorogenic acid B, isochlorogenic acid C and eleutheroside B were identified as potential Q-Markers of Acanthopanacis Cortex for “same treatment for different diseases” in RA and OP.

R282.6

国家自然科学基金青年科学基金项目（82404859）；江苏省植物分类与资源保护利用重点实验室开放基金项目（JSPKLB202408）；江苏省药品监督管理局药品监管科学科研计划项目（202421）