三黄汤降糖降脂活性成分的多靶标亲和超滤质谱研究

Study on Hypoglycemic and Hypolipidemic Compounds in Sanhuang Decoction by Multi-target Affinity Ultrafiltration LC-MS

三黄汤是传统的中药复方,临床上被用于治疗糖尿病和肥胖已有上千年的历史,但三黄汤中具有显著降血糖和降血脂作用的活性成分及其作用机制仍不清楚。为此,本研究首先测试三黄汤对α-葡萄糖苷酶(α-Glu)和脂肪酶(LP)的抑制活性,然后分别以α-Glu、α-淀粉酶(α-Amy)和LP这3个靶酶为靶标,采用多靶标亲和超滤质谱技术从三黄汤中筛选出24、17和25种分别能够与α-Glu、α-Amy和LP结合的潜在活性配体,其中,表儿茶素-3-O-没食子酸酯等多个成分具有显著的降糖和降脂活性。随后,对筛选出的部分代表性活性成分进行分子对接验证。本研究结合3种糖脂代谢相关药物靶标,采用多靶标亲和超滤质谱技术探究三黄汤复方中潜在的降糖降脂活性成分,并据此构建多成分多靶标作用网络图,可为阐明降糖降脂活性物质基础和防治肥胖和糖尿病的临床应用提供理论依据。

Sanhuang decoction,a traditional Chinese medicine prescription,was firstly described in Golden Chamber by Zhang Zhongjing in the Han Dynasty,and has been used clinically for thousands of years to treat diabetes and obesity.However,the specific bioactive compounds being responsible for the hypoglycemic and hypolipidemic effects of Sanhuang decoction and the underlying therapeutic mechanism remain elusive,and further research efforts are in great need.Hence,the aim of this study was to discover the potential hypoglycemic and hypolipidemic phytochemicals in Sanhuang decoction and to explore their potential mechanisms of action.Here,the in vitroα-glucosidase(α-Glu)and lipase(LP)inhibitory activities of Sanshuang decoction were determined,and the results showed that Sanhuang decoction possesses significantα-Glu and LP inhibitory activities in vitro.Then,24,17 and 25 potential bioactive ligands in Sanhuang decoction are successfully captured by combining affinity ultrafiltration liquid chromatography-mass spectrometry with three drug targets,namelyα-Glu,α-amylase(α-Amy)and LP,respectively.These potential bioactive ingredients fished out from Sanhuang decoction exhibit various binding capacities toα-Glu,α-Amy and LP,respectively.Thereof,epicatechin-3-O-gallate possesses significant hypoglycemic and hypolipidemic effects.Finally,some representative active ligands were further verified with molecular docking assay,and the molecular docking results are consistent with those of affinity ultrafiltration.Furthermore,in addition to hydrogen bond(H-bond),there are various other interaction forces between these potential bioactive ingredients and their corresponding target enzymes,such as Pi-Sigma,Pi-Pi T-shaped and Pi-Alkyl interactions.In this study,the potential hypoglycemic and hypolipidemic constituents in Sanshuang decoction were explored and identified by using affinity ultrafiltration liquid chromatography-mass spectrometry with three drug targets tightly correlating to the metabolisms of blood sugar and lipids in the human body,and the effective material basis of Sanshuang decoction can thus be preliminarily clarified.In this way,the interaction network among these bioactive compounds and three drug targets were constructed,thus providing theoretical support for the clinical applications of Sanhuang decoction for the treatments of obesity and diabetes.