目的 采用精准、可量化的方法筛选蜂胶影响PXR/CYP3A4通路调控血脂的质量标志物。方法 采用人结肠癌LS174T细胞给予一定浓度的咪达唑仑注射液，同时给予不同浓度的蜂胶已知成分对照品溶液，孵育后提取样品，采用UHPLC-MS法测定生成的1'-羟基咪达唑仑含量，根据测定结果筛选出对PXR/CYP3A4通路有显著调控作用的成分，以这些成分为指标，建立同时定量方法，根据结果初步确定蜂胶中影响PXR/CYP3A4通路调控血脂代谢的质量标志物。结果 所评价的蜂胶成分中，白杨素、高良姜素、异绿原酸A、槲皮素、咖啡酸苯乙酯均能够显著提高或降低1'-羟基咪达唑仑的量（与对照组和阳性对照组比较），提示上述成分能够显著影响PXR/CYP3A4表达；UHPLC-MS-MS含量测定结果显示，蜂胶中上述成分除异绿原酸A和乔松素外，均具有适宜的含量以达到显效。结论 白杨素、高良姜素、咖啡酸苯乙酯和槲皮素可能是蜂胶影响PXR/CYP3A4通路调控血脂的质量标志物，且均具有抑制相关靶标表达，进而调节血脂水平的活性；同时，本研究所用质量标志物筛选、评价方法快速、高效、可量化，能够适应基于PXR/CYP3A4的大通量活性成分筛选研究。

Objective To screen and evaluate PXR/CYP3A4-induced lipid-regulating quality marker in propolis with precise and quantitative method. Methods The LS174T cell was given certain amount of midazolam injection, along with different dosage of known components found in propolis, after incubation and extraction, the samples were determined for 1'-OH-midazolam, and each compound was evaluated to discover the PXR/CYP3A4 pathway regulatory activity according to the results; Then, compounds selected were used as indexes for UHPLC-MS-MS content determination, and their own values were regarded as a preliminary step of confirming PXR/CYP3A4-induced lipid-regulating quality markers of propolis. Results In all components tested, chrysin, galangin, heterochlorogenic acid A, quercetin, and caffeic acid phenethylester significantly affected the 1'-OH-midazolam yield compared with blank and positive control, indicating their obvious influence on PXR/CYP3A4 expression; The UHPLC-MS-MS determination showed that except galangin, heterochlorogenic acid A, and quercetin, all the other compounds had adequate content in propolis to take effect. Conclusion Chrysin, galangin, caffeic acid phenethylester, and quercetin were probably defined as PXR/CYP3A4-induced lipid-regulating quality marker in propolis, which inhibited the expression of such targets to down-regulate blood lipid level; Additionally, the method used for quality marker screening and evaluation in this study was fast, effective and quantitative, and capable of carrying out high throughput active component screening for PXR/CYP3A4 regulatory activities.

R285.5

国家自然科学基金资助项目（81903761）；国家重大科技专项（2016ZX09101076）；广东省科技计划项目：创新中药研发平台建设（2017A070701017）；广东省中医药局课题（20192006）