2型糖尿病气阴两虚证大鼠尿液代谢组学研究

Urine metabonomics in type 2 diabetes rats with qi yin deficiency syndrome

目的 通过尿液非靶向代谢组学方法,从代谢差异标志物及代谢通路方面,探讨成年SD大鼠2型糖尿病气阴两虚证的中医证候内在的生物物质基础。方法 将SD大鼠随机分为正常组和2型糖尿病(diabetes mellitus type 2,T2DM)气阴两虚证组,采用高脂高糖饲料饲喂、小剂量链脲佐菌素(streptozocin, STZ)35 mg/kg腹腔注射结合青皮附子散颗粒剂灌胃的方法建立T2DM气阴两虚证模型。利用液质联用(Liquid Chromatography-Mass Spectrometry, LC-MS)技术进行尿液非靶代谢组学分析,运用主成分分析(Principal Component Analysis, PCA)和偏最小二乘法判别分析(Partial Least Squares Discriminant Analysis, PLS-DA),KEGG数据库、HMDB数据库、LIPIDMaps数据库和液相色谱与串联质谱联用(liquid chromatography tandem mass spectrometry, LC-MS/MS)等鉴定潜在的生物标志物进行通路富集分析。结果 发现与糖尿病发生相关的代谢物变化涉及碳水化合物代谢9条,脂质代谢12条,氨基酸代谢9条,能量代谢2条。结论 糖尿病气阴两虚证生物模型存在碳水化合物代谢、脂质代谢、氨基酸代谢、生物能量代谢紊乱,代谢组学研究方法有助于揭示中医证候的微观物质基础。

Objective To explore the intrinsic biomaterial basis of the TCM syndrome of type 2 diabetes in adult SD rats through differential metabolic markers and metabolic pathways using urine untargeted metabolomics.Methods SD rats were randomly divided into normal and T2DM groups with deficiency of both qi and yin.The T2DM group with deficiency of both qi and yin was established by feeding a high fat and sugar diet combined with intraperitoneal injection of a small dose of streptozotocin(35 mg/kg)and intragastric administration of Qingpi Fuzi powder granules.Untargeted metabonomic analysis of urine was performed by liquid chromatography-tandem mass spectrometry.Potential biomarkers and pathway enrichment were identified by principal component analysis,partial least squares discriminant analysis,KEGG database,HMDB database,LIPIDMaps database,and MS/MS tandem ion fragment information.Results Metabolite changes related to diabetes were found to involve nine carbohydrate,twelve lipid,nine amino acid,and two energy metabolic pathways.Conclusions There are carbohydrate,lipid,amino acid,and bioenergy metabolism disorders in the biological model of qi yin deficiency in diabetes.Metabonomics are helpful to reveal the material basis of TCM syndromes.