基于脂质效应分子筛选泽泻三萜调节脂代谢紊乱的质量评价标志物

Alismatis Rhizoma (AR) is a well-known Chinese medicine and has been commonly used for the treatment of various diseases, including dysuria and hyperlipidemia. Our previous study demonstrated that triterpenes from the AR potentially regulated to the lipid disorders. However, alisol B 23-acetate, one of the main triterpenes in AR, was used as the marker of quality control for AR in 2015 Chinese Pharmacopoeia, while it can not reflect the intrinsic quality of AR on lipid-lowering effect. In addition, it was impossible to carry out quality control on AR from the perspective of lipid-lowering ingredients. In this study, a specific characteristic chemical profile for systematic identification and specific characterization of phospholipid, cholesteryl ester and triacylglycerol in plasma will be established by integrated strategy combining QTOF-MS/MS and QTRAP-MS/MS, which will be used for high-throughput screening of lipid effect molecules regulated by triterpenes on normal mice and high-fat mice. Afterwards, a characteristic-nonlinear effect model between lipid effect molecules and triterpenes will be constructed for screening bioactive triterpenes as the quality control marker of AR by support vector machine. At last, a scientific, reasonable and effective quality evaluation method will be established by the bioactive triterpenes. In conclusion, this study will provide a new way for exploring the effective control of the intrinsic quality of Chinese medicines with lipid-lowering effect.

泽泻为临床常用中药，具有利水渗湿、化浊降脂之功效。前期研究表明：泽泻中的三萜是调节脂代谢紊乱的药效成分。2015版《中国药典》仅以三萜23-乙酰泽泻醇B作为泽泻的含量测定指标，难以准确评价其降脂的内在品质，无法从降脂活性成分的角度对泽泻进行质量控制。本项目拟在前期研究基础上，通过组合运用QTOF-MS/MS和QTRAP-MS/MS两种质谱技术，建立全面表征和靶标性鉴定血清磷脂、胆固醇酯和三酰甘油酯三大类脂质成分的特异性表征谱分析方法，用于高通量筛选正常小鼠和高脂小鼠血清中，共同被泽泻三萜调节的药物敏感性脂类成分——脂质效应分子；结合支持向量机的机器学习方法，拟合脂质效应分子与三萜成分之间的“特征-非线性效应”模型，筛选能够客观反映泽泻降脂作用的质量控制标志物组分，为泽泻建立一种科学、合理和有效的质量评价方法。该研究为探索有效控制降脂中药的内在质量，提供了一种新的模式和经验借鉴。

本项目组合运用UHPLC-QTOF-MS/MS和UPLC-QTRAP-MS/MS技术建立了血清脂质特异性表征谱分析方法，共鉴定出216个磷脂和55个胆固醇酯，可以用高通量靶向分析血清中的脂质成分。并将该方法首次用于筛选泽泻三萜调节脂代谢紊乱的脂质效应分子，在高脂小鼠血清中发现了61代谢紊乱的脂质成分，泽泻三萜对其中的47个脂质成分具有调节作用。在此基础上，运用SVM模型首次构建了脂质效应分子和泽泻三萜成分间的“特征-非线性效应”模型，筛选出了9个评价泽泻降脂作用的质量控制三萜标志物组分。此外，为了阐明泽泻三萜的降脂作用机理，本项目将脂质组学和代谢组学的研究方法和网络药理学进行了整合，首次发现泽泻三萜可调节脂代谢、能量代谢和氨基酸代谢，并通上调ALB、TNF、IL1B、MMP9、PPARA和PPARG这6个靶点起到降脂作用，其中PPAR信号通路与泽泻降脂作用最为相关。通过以上研究，本项目筛选出了泽泻降脂的质量控制标志物，并阐明了泽泻三萜调节脂代谢紊乱的作用机制。为有效控制降脂中药的内在质量、复杂中药组分药效物质筛选提供了成功经验。