非靶向尿液代谢组学方法研究五味子治疗精尿病并发症作用机制

摘要

采用超高效液相色谱-串联四级杆飞行时间质谱联用技术(Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry,UPLC/Q-TOF-MS),结合主成分分析技术(Principal component analysis,PCA),进行了五味子治疗糖尿病并发症大鼠尿液代谢组学研究,对五味子治疗糖尿病并发症大鼠后尿液生物标志物变化进行了考察.采用UPLC/Q-TOF-MS方法分析了健康组、糖尿病并发症模型组和五味子给药组的大鼠尿液,采用偏最小二乘法-判别分析(Partial least squares discriminant analysis,PLS-DA)和正交偏最小二乘法-判别分析(Orthogonal partial least squares discriminant analysis,OPLS-DA)载荷图筛选,通过分析对各组分离贡献较大(VIP>1,P°0.05)化合物的串联质谱数据,再经Human Metabolome Data-base(HMDB)等数据库检索,进行质谱信息匹配,尿液中共鉴定出28种潜在生物标记物,其中正谱13种,负谱15种.这些生物标记物主要影响戊糖和葡糖醛酸相互转化通路、核黄素代谢、泛酸和CoA合成、精氨酸和脯氨酸代谢、肠内菌代谢、嘌吟代谢、Vc代谢胆酸合成、色氨酸代谢等通路,五味子从能量代谢、氨基酸代谢、肠内菌代谢、脂类代谢等各个角度发挥治疗糖尿病并发症的作用.从各个通路的相关生物功能分析,五味子治疗糖尿病肾病的作用较强,此外还具有降脂、抗氧化的功效.%An untargeted urinary metabonomics method based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry ( UPLC-Q-TOF-MS ) has been established to investigate the mechanism of Schisandra chinensis in treating diabetes and its complications. The urinary biomarkers related to the therapeutic effects of Schisandra chinensis on the diabetes rats were analyzed. In urine, 28 kinds of endogenous metabolites were identified as potential biomarkers, including 13 endogenous metabolites in positive ion mode, 15 endogenous metabolites in negative ion mode, and hippuric acid detected both in positive and negative ion modes. The results revealed that Schisandra chinensis mainly affected the pathways of pentose and glucuronate interconversions, riboflavin metabolism, pantothenate and CoA biosynthesis, arginine and proline metabolism, intestinal bacteria metabolism, ascorbate and aldarate metabolism and tryptophan metabolism in diabetic rats. Combined with biological analysis of these pathways, the therapeutic mechanism of Schisandra chinensis on diabetes and its complications was verified. Based on the biological function of each pathway, the effect of Schisandra chinensis on diabetic nephropathy is stronger. Moreover, it also has the effects of protecting liver, decreasing fat and antioxidant activity.