Table_2_Mechanism of Paeoniae Radix Alba in the Treatment of Non-alcoholic Fatty Liver Disease Based on Sequential Metabolites Identification Approach, Network Pharmacology, and Binding Affinity Measurement.XLSX

Screening functional food ingredients (FFI) from medicinal and edible plants (MEP) has still remained a great challenge due to the complexity of MEP and its obscure function mechanisms. Herein, an integrated strategy based on sequential metabolites identification approach, network pharmacology, molecular docking, and surface plasmon resonance (SPR) analysis was proposed for quickly identifying the active constituents in MEP. First, the sequential biotransformation process of MEP, including intestinal absorption and metabolism, and hepatic metabolism, was investigated by oral gavage, and intestinal perfusion with venous sampling method. Then the blood samples were analyzed by UPLC-Q Exactive Orbitrap HRMS. Second, the network pharmacology approach was used to explore the potential targets and possible mechanisms of the in vivo metabolites of MEP. Third, molecular docking and SPR approaches were used to verify the specific interactions between protein targets and representative ingredients. The proposed integrated strategy was successfully used to explore the heptoprotective components and the underlying molecular mechanism of Paeoniae Radix Alba (PRA). A total of 44 compounds were identified in blood samples, including 17 porotypes and 27 metabolites. The associated metabolic pathways were oxidation, methylation, sulfation, and glucuronidation. After further screening, 31 bioactive candidates and 377 related targets were obtained. In addition, the bioactive components contained in PRA may have therapeutic potentials for non-alcoholic fatty liver disease (NAFLD). The above results demonstrated the proposed strategy may provide a feasible tool for screening FFI and elaborating the complex function mechanisms of MEP.

从药食同源植物（medicinal and edible plants, MEP）中筛选功能性食品原料（functional food ingredients, FFI）仍面临巨大挑战，这源于药食同源植物本身的复杂性及其作用机制的模糊性。为此，本研究提出了一种基于序列代谢物鉴定法、网络药理学、分子对接以及表面等离子体共振（surface plasmon resonance, SPR）分析的整合策略，用于快速鉴定药食同源植物中的活性成分。首先，采用灌胃给药结合静脉采血法的肠灌注实验，考察药食同源植物的序列生物转化过程——包括肠道吸收、代谢及肝脏代谢。随后采用超高效液相色谱-静电场轨道阱高分辨质谱（UPLC-Q Exactive Orbitrap HRMS）对血液样本进行分析。其次，借助网络药理学方法，探究药食同源植物体内代谢物的潜在作用靶点与潜在作用机制。最后，通过分子对接与表面等离子体共振技术，验证蛋白靶点与代表性成分之间的特异性相互作用。本研究将所提出的整合策略成功应用于白芍（Paeoniae Radix Alba, PRA）的保肝活性成分及其潜在分子机制探究。最终在血液样本中共鉴定出44种化合物，其中包括17种原型成分与27种代谢产物。相关代谢途径涵盖氧化、甲基化、硫酸化及葡萄糖醛酸化反应。经进一步筛选，最终得到31个活性候选成分与377个相关作用靶点。此外，白芍中含有的活性成分对非酒精性脂肪性肝病（non-alcoholic fatty liver disease, NAFLD）具有潜在治疗价值。上述研究结果表明，本研究提出的整合策略可为功能性食品原料的筛选以及药食同源植物复杂作用机制的阐释提供可行的研究工具。