目的 基于特征图谱、网络药理学及定量分析方法，分析预测牡荆叶Viticis Negundo Folium标准汤剂的潜在质量标志物（quality markers，Q-Marker），构建牡荆叶标准汤剂的多维度质量评价体系。方法 采用UPLC建立牡荆叶标准汤剂的特征图谱，色谱条件：色谱柱为Cortecs C₁₈柱（100 mm×2.1 mm，2.7 μm），流动相为乙腈-0.1%磷酸水溶液，梯度洗脱：0～9 min，3%～16%乙腈；9～21 min，16%～20%乙腈；21～22 min，20%～65%乙腈；22～23 min，65%乙腈；23～29 min，65%～95%乙腈；检测波长270 nm，柱温35 ℃，体积流量0.3 mL/min，进样体积2 μL。通过网络药理学预测候选Q-Marker的关键靶点和关键通路，构建“候选Q-Marker-关键靶点-关键通路”网络。对牡荆叶候选Q-Marker进行定量分析，明确牡荆叶候选Q-Marker在饮片-标准汤剂的量值传递规律。结果 建立了牡荆叶标准汤剂的特征图谱，共标定出9个特征峰，通过对照品确认其中4个色谱峰（峰1、3、5、8），分别为对羟基苯甲酸、异荭草苷、异牡荆苷、芹菜素-7-O-β-D-葡萄糖醛酸苷（apigenin-7-O-β-D-glucuronide，AG）。经网络药理学分析，牡荆叶标准汤剂中对羟基苯甲酸、异荭草苷、异牡荆苷、AG，可能作用于基质金属蛋白酶2（matrix metalloproteinase 2，MMP2）、v-Rel禽网状内皮组织增殖病毒癌基因同源物A（v-Rel avian reticuloendotheliosis virus oncogene homolog A，RELA）、组蛋白去乙酰化酶1（histone deacetylase 1，HDAC1）等关键靶点，调控细胞衰老、晚期糖基化终末产物-受体（advanced glycation end products-receptor for advanced glycation end products signaling pathway，AGE-RAGE）、磷脂酰肌醇3-激酶-蛋白激酶B（phosphatidylinositol 3-kinase-protein kinase B，PI3K-Akt）等关键通路发挥治疗支气管炎的作用。对这4个成分进行定量分析，15批牡荆叶标准汤剂中的平均质量分数分别为18.98、9.13、4.72、1.64 mg/g，饮片中的平均质量分数分别为4.98、11.44、2.38、2.60 mg/g，从饮片到标准汤剂的平均转移率分别为118.83%、24.61%、54.75%、19.81%。初步确定对羟基苯甲酸、异荭草苷、异牡荆苷、AG为牡荆叶治疗支气管炎的Q-Marker。结论 建立的质量评价方法准确可靠，量值传递均一稳定，结合网络药理学筛选的潜在Q-Marker可为牡荆叶标准汤剂的质量控制提供依据，为后续阐明其药效物质基础的作用机制提供支撑。

Objective To analyze and predict the quality markers (Q-Markers) of Mujingye (Viticis Negundo Folium, VNF) standard decoction based on characteristic chromatogram, network pharmacology, and quantitative analysis, and to construct a multi-dimensional quality evaluation system for VNF standard decoction. Methods UPLC was used to establish the characteristic chromatogram of VNF standard decoction. The chromatographic conditions were as follows: Cortecs C₁₈ column (100 mm×2.1 mm, 2.7 μm); the mobile phase consisted of acetonitrile (A) and 0.1% phosphoric acid (B), using gradient elution (0—9 min, 3%—16% A; 9—21 min, 16%—20% A; 21—22 min, 20%—65% A; 22—23 min, 65% A; 23—29 min, 65%—95% A); the detection wavelength was 270 nm; the column temperature was 35 ℃; the flow rate was 0.3 mL/min; and the injection volume was 2 μL. Network pharmacology was employed to obtain the key targets and key pathways of candidate Q-Markers, and the “candidate Q-Marker-key target-key pathway” network was constructed. Quantitative analysis was performed on the candidate Q-Markers of VNF to clarify the rules of quantity transfer of candidate Q-Markers from decoction pieces to standard decoction. Results The characteristic chromatogram of VNF standard decoction was established, and nine characteristic peaks were marked. Among them, four chromatographic peaks (peaks 1, 3, 5, and 8) were identified by reference substances as p-hydroxybenzoic acid, isoorientin, isovitexin, and apigenin-7-O-β-D-glucuronide (AG). Network pharmacology analysis revealed that p-hydroxybenzoic acid, isoorientin, isovitexin, and AG in VNF standard decoction could act on key targets such as matrix metalloproteinase 2 (MMP2), v-Rel avian reticuloendotheliosis virus oncogene homolog A(RELA), and histone deacetylase 1 (HDAC1), and regulate key pathways including cellular senescence, advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signaling pathway, and phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) signaling pathway to exert therapeutic effects against bronchitis. Quantitative analysis of these four components showed that their average contents in 15 batches of VNF standard decoction were 18.98, 9.13, 4.72, 1.64 mg/g, respectively, while their average contents in the decoction pieces were 4.98, 11.44, 2.38, 2.60 mg/g, respectively. The average transfer rates from decoction pieces to standard decoction were 118.83%, 24.61%, 54.75%, 19.81%, respectively. p-Hydroxybenzoic acid, isoorientin, isovitexin, and AG were preliminarily identified as the Q-Markers of VNF for treating bronchitis. Conclusion The established quality evaluation method is accurate and reliable, with uniform and stable quantity transfer. Combined with the Q-Markers screened by network pharmacology, it can provide a basis for the quality control of VNF standard decoction and support for the subsequent elucidation of the mechanism of action of its pharmacodynamic material basis.

R283.6

广西科技重大专项（桂科AA22096019）；河池市中央引导地方科技发展专项（河科ZY230303）