Polysaccharides are widely present in herbs with multiple activities, especially immunity regulation and metabolic benefits for metabolic disorders. However, the underlying mechanisms are not well understood. Functional metabolomics is increasingly used to investigate systemic effects on the host by identifying metabolites with particular functions. This study explores the mechanisms underlying the metabolic benefits of Astragalus polysaccharides (APS) by adopting a functional metabolomics strategy. The effects of APS were determined in eight-week high-fat diet (HFD)-fed obese mice. Then, gas chromatography–time-of-flight mass spectrometry (GC–TOFMS)-based untargeted metabolomics was performed for an analysis of serum and liver tissues, and liquid chromatography–tandem mass spectrometry (LC–MS/MS)-based targeted metabolomics was performed. The potential functions of the metabolites were tested with in vitro and in vivo models of metabolic disorders. Our results first confirmed the metabolic benefits of APS in obese mice. Then, metabolomics analysis revealed that APS supplementation reversed the HFD-induced metabolic changes, and identified 2-hydroxybutyric acid (2-HB) as a potential functional metabolite for APS activity that was significantly decreased by a HFD and reversed by APS. Further study indicated that 2-HB inhibited oleic acid (OA)-induced triglyceride (TG) accumulation. It was also found to stimulate the expression of proteins in lipid degradation in hepatocytes and TG lipolysis in 3T3-L1 cells. Moreover, it was found to reduce serum TG and regulate the proteins involved in lipid degradation in high-fat and high-sucrose (HFHS)-fed mice. In conclusion, our study demonstrates that the metabolic benefits of APS are at least partially due to 2-HB generation, which modulated lipid metabolism both in vitro and in vivo. Our results also highlight that functional metabolomics is practical for investigating the mechanism underlying the systemic benefits of plant polysaccharides.

多糖广泛存在于具有多种活性的草药中，尤其是对代谢紊乱的免疫调节和代谢益处。然而，潜在的机制尚不清楚。功能代谢组学越来越多地用于通过识别具有特定功能的代谢物来研究对宿主的全身影响。本研究探讨了黄芪代谢益处的潜在机制多糖（APS）通过采用功能代谢组学策略。APS 的影响是在八周高脂饮食 (HFD) 喂养的肥胖小鼠中确定的。然后，进行基于气相色谱-飞行时间质谱 (GC-TOFMS) 的非靶向代谢组学分析血清和肝组织，以及基于液相色谱-串联质谱 (LC-MS/MS) 的靶向代谢组学进行了。通过体外和体内测试代谢物的潜在功能代谢紊乱模型。我们的研究结果首先证实了 APS 在肥胖小鼠中的代谢益处。然后，代谢组学分析显示，补充 APS 可逆转 HFD 诱导的代谢变化，并确定 2-羟基丁酸 (2-HB) 是 APS 活性的潜在功能代谢物，HFD 显着降低该活性并被 APS 逆转。进一步的研究表明，2-HB 抑制油酸 (OA) 诱导的甘油三酯 (TG) 积累。还发现它可以刺激肝细胞脂质降解中蛋白质的表达和 3T3-L1 细胞中的 TG 脂肪分解。此外，在高脂肪和高蔗糖 (HFHS) 喂养的小鼠中，发现它可以降低血清 TG 并调节参与脂质降解的蛋白质。综上所述，体外和体内。我们的研究结果还强调，功能代谢组学对于研究植物多糖全身益处的机制是实用的。