目的基于“性-效-物”三元论研究原则，预测桐桔梗质量标志物（Q-Marker），阐明其苦味药性与抗炎、祛痰功效的物质基础及作用机制。方法采用HPLC-QTOF-MS对桐桔梗进行物质组解析；通过味觉受体分子对接结合电子舌技术探究其苦味物质基础；采用脂多糖（LPS）诱导RAW 264.7细胞炎症模型、表皮生长因子（EGF）诱导的NCI-H292细胞黏液高分泌模型，结合核因子（NF）-κB报告基因、β2肾上腺素受体（ADRB2）转染细胞系、5-脂氧合酶（5-LOX）及环氧合酶2（COX-2）抑制剂筛选体系，考察桐桔梗主要化学成分的抗炎、祛痰作用及作用机制。结果从桐桔梗中共鉴定出154个化合物，其中皂苷类77个；分子对接及电子舌结果表明，三萜皂苷类成分为其主要苦味物质基础。结合前期研究与本实验结果，桔梗皂苷D（PD）、桔梗皂苷D₃（PD₃）、桔梗皂苷E（PE）、去芹糖桔梗皂苷D（DPD）、远志皂苷D（PGD）、远志皂苷D₂（PGD₂）、桔梗酸A（PCA）、去芹糖桔梗皂苷D₃（DPD₃）、去芹糖桔梗皂苷E（DPE），9个入血原型三萜皂苷均可显著降低LPS诱导的RAW 264.7细胞上清液中NO及多种炎症因子水平；桐桔梗总提物、不同极性部位及PD、DPD、PD₃、PE、PCA、DPD₃可显著抑制EGF诱导NCI-H292细胞中活性氧（ROS）水平（P＜0.05、0.01、0.001）。桐桔梗总提物、不同极性部位及PD、DPD、PE、PGD₂、PCA、DPD₃、DPE均可显著抑制EGF诱导NCI-H292细胞中黏蛋白5AC（MUC5AC）表达（P＜0.05、0.01、0.001）。其作用机制涉及抑制NF-κB通路转录活性、COX-2和5-LOX酶活性，以及激活ADRB2信号通路。结论基于“性-效-物”三元论研究路径，初步阐明桐桔梗药性（味）及药效的物质基础，筛选得到9个三萜皂苷，作为基于体外“性-效-物”关联的候选Q-Marker，为桐桔梗质量标准提升提供参考。

Objective To predict quality markers (Q-Marker) of Platycodonis Radix based on the “property-response-component” triad research principle, and to elucidate the material basis and mechanisms underlying its bitter taste and anti-inflammatory and expectorant effects. Methods The chemical profile of Platycodonis Radix was analyzed using HPLC-QTOF-MS; the molecular basis of its bitterness was investigated through molecular docking with taste receptors combined with electronic tongue technology. The antiinflammatory and expectorant activities and their mechanisms were evaluated using an LPS-induced RAW 264.7 cell inflammation model and an EGF-induced NCI-H292 cell mucus hypersecretion model, in combination with NF-κB reporter gene assays, β2- adrenergic receptor (ADRB2) stably transfected cell line stably transfected cell lines, and inhibition systems for 5-lipoxygenase (5- LOX) and cyclooxygenase-2 (COX-2). Results A total of 154 compounds were identified from Platycodonis Radix, including 77 saponins. Molecular docking and electronic tongue analyses indicated that triterpene saponins are the primary contributors to its bitter taste. Based on previous studies and current findings, nine prototype triterpene saponins—platycodin D (PD), platycodin D₃ (PD₃), platycoside E (PE), deapioplatycodin D (DPD), polygalacin D (PGD), polygalacin D₂ (PGD₂), platyconic acid A (PCA), deapioplatycodin D₃ (DPD₃), and deapiplatycoside E (DPE)—significantly reduced NO levels and multiple inflammatory cytokines in the supernatant of LPS-stimulated RAW 264.7 cells. Total extract, different polarity fractions, and individual compounds PD, DPD, PD₃, PE, PCA, and DPD₃ significantly suppressed ROS production in EGF-induced NCI-H292 cells (P < 0.05, 0.01, 0.001). Moreover, total extract, different polarity fractions, and individual compounds PD, DPD, PE, PGD₂, PCA, DPD₃, and DPE significantly reduced MUC5AC expression in EGF-induced NCI-H292 cells (P < 0.05, 0.01, 0.001). The underlying mechanisms involved inhibition of NF-κB pathway transcriptional activity and COX-2 and 5-LOX enzyme activities, as well as activation of the ADRB2 signaling pathway. Conclusion This study, guided by the “property-response-component” triad approach, has preliminarily clarified the material basis of the pharmacological properties (taste) and effects of Platycodonis Radix, identifying nine triterpene saponins as candidate Q-Marker based on in vitro property-response-component associations, providing valuable insights for improving the quality standards of this herb.

R285.5

国家科技重大专项-“四大慢病重大专项”（2025ZD0549700）；国家自然科学基金资助项目（ U21A20406）