目的 采用超高效液相色谱-四级杆-飞行时间质谱（ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry，UPLC-Q-TOF-MS/MS）结合特征分子网络（feature-based molecular networking，FBMN）技术，对畲药树参Dendropanax dentiger醇提物化学成分进行快速鉴定，并对其抗炎活性进行初步评价。方法 ACQUITY UPLC HSS T3色谱柱（100 mm×2.1 mm，1.8 μm）；以0.1%甲酸水（A）-乙腈（B）为流动相梯度洗脱，在正、负离子模式下采集质谱数据，并在全球天然产物社会分子网络（global natural products social molecular networking，GNPS）网站（http://gnps.ucsd.edu）上构建FBMN。基于SIRIUS 6.2.2软件及GNPS数据库匹配结果，结合对照品比对、保留时间、特征碎片、文献及PubChem、MassBank数据库信息，并结合FBMN的结构聚类特性，快速鉴定树参醇提物的化学成分并总结其裂解规律；采用脂多糖（lipopolysaccharide，LPS）诱导的RAW264.7巨噬细胞炎症模型，结合CCK-8、Western blotting及免疫荧光实验，评价其抗炎效果。结果 共鉴定出树参中59种化学成分，包括酚酸类18个、木脂素类11个、黄酮及其苷类8个、香豆素6个、环肽4个、苯乙醇苷3个、脂肪酸及其衍生物4个、环烯醚萜2个、糖苷2个及生物碱1个。其中，19个潜在未报道化合物首次从树参中分析得到。基于FBMN，成功识别异绿原酸A、B、C等同分异构体，并进一步推测出其甲基化衍生物。细胞实验结果表明，树参醇提物可显著抑制LPS诱导的炎症反应，降低白细胞介素-1β（interleukin-1β，IL-1β）和肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）的表达，促进白细胞介素-10（interleukin-10，IL-10）分泌，并抑制核因子-κB亚基p65（nuclear factor-kappa-B p65，NF-κB p65）的核转位。结论 FBMN弥补了传统分子网络在鉴定同分异构体方面的不足，UPLC-Q-TOF-MS/MS联合FBMN技术可实现树参复杂成分的快速、高效鉴定，并首次发现19个潜在未报道化合物。树参提取物具有良好抗炎活性，为其后续药效物质研究和开发应用提供实验依据。

Objective To rapidly identify the chemical constituents of the ethanol extract of Dendropanax dentiger, a She ethnic medicine, using ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) and feature-based molecular networking(FBMN), and preliminarily evaluate its anti-inflammatory activity. Methods Chromatographic separation was performed on an ACQUITY UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm) using a gradient elution with 0.1% formic acid in water (A) and acetonitrile (B) as the mobile phases. MS data were acquired in both positive and negative ion modes, and FBMN was constructed on the global natural products social molecular networking (GNPS) platform (http://gnps.ucsd.edu). Based on the matching results of the SIRIUS 6.2.2 software and the GNPS database, combined with the reference standards, retention time, feature fragments, literature, and information from the PubChem and MassBank databases, and in combination with the structural clustering characteristics of FBMN, the chemical composition of the alcohol extract of D. dentiger was rapidly identified and its lysis rules were summarized. In vitro anti-inflammatory effects were evaluated using a lipopolysaccharide (LPS)-induced RAW264.7 macrophage inflammation model, with CCK-8, Western blotting, and immunofluorescence analyses. Results A total of 59 compounds were identified, including 18 phenolic acids, 11 lignans, eight flavonoids and flavonoid glycosides, six coumarins, four cyclic peptides, three phenylethanoid glycosides, four fatty acids and derivatives, two iridoids, two glycosides, and one alkaloid. Among them, 19 potentially unreported compounds were identified from D. dentiger for the first time. Three positional isomers—isochlorogenic acid A, B, and C were successfully identified via FBMN, and a methylated derivative was further proposed. The results of cell experiments demonstrated that the ethanol extract of D. dentiger significantly suppressed LPS-induced inflammatory responses in RAW264.7 cells by downregulating interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) expression, upregulating IL-10 secretion, and inhibiting nuclear translocation of nuclear factor-kappa-B p65(NF-κB p65) . Conclusion FBMN effectively overcomes the limitations of traditional molecular networking in isomer identification. The integration of UPLC-Q-TOF-MS/MS with FBMN enables rapid and efficient identification of the complex components in D. dentiger, leading to the first identification of 19 potentially unreported compounds. The ethanol extract of D. dentiger exhibits promising anti-inflammatory activity, providing an experimental basis for further research and development application of its active substances.

R284.1

浙江省中医药科技计划项目（2024ZR205）；浙江省中医药管理局共建科技计划项目（GZY-ZJ-KJ-23098）；丽水市公益性技术应用研究项目（2024GYX36）；丽水市自筹类公益性技术应用研究项目（2022SJZC037）