基于代谢组学探讨玄参-苍术联合二甲双胍治疗2型糖尿病大鼠作用机制

Approach to mechanism of scrophularia-atractylodes and metformin on type 2 diabetes mellitus based on metabolomics

目的:采用代谢组学方法分析玄参-苍术联合二甲双胍治疗2型糖尿病大鼠的作用机制。方法:采用高脂饮食联合链脲佐菌素(streptozocin,STZ)建立2型糖尿病大鼠模型,分为空白组、模型组、阳性组和治疗组。治疗后6周,检测各组大鼠体质量、血糖、丙氨酸氨基转移酶、空腹血清胰岛素、核转录因子(NF-κB)、大鼠隐热蛋白(NLRP3)等指标水平,初步评价玄参-苍术联合二甲双胍对2型糖尿病大鼠的治疗作用。进一步采用UPLC-MS/MC非靶向代谢组学揭示各组之间的代谢谱,筛选差异代谢物并进行鉴别。结果:与阳性组比较,治疗组改善临床症状及降低餐后高血糖的作用更为明显(P﹤0.05),且可显著降低NF-κB和NLRP3水平(P﹤0.01)。通过代谢组学筛选出36种差异代谢物作为玄参-苍术联合二甲双胍治疗糖尿病作用机制的相关血清潜在生物标志物。D-谷氨酰胺和D-谷氨酸是玄参-苍术联合二甲双胍治疗糖尿病涉及的相关的代谢通路。结论:玄参-苍术联合二甲双胍治疗糖尿病有效,推测其机制是通过调节D-谷氨酰胺和D-谷氨酸代谢通路基因的表达以维持“谷氨酰胺-谷氨酸”动态平衡,抑制NF-κB/NLRP3通路激活,进而提高机体对胰岛素敏感性、调节糖脂代谢紊乱、减轻炎症反应。

Objective:To approach the mechanism of scrophularia-atractylodes and metformin on type 2 diabetes mellitus by metabolomics.Methods:The model of type 2 diabetic rat was established by high-fat diet and streptozocin.The type 2 diabetic rats were divided into blank group,model group,positive group and treatment group.After 6 weeks of treatment,body weight,fasting blood glucose,alanine transaminase,fasting serum insulin,nuclear transcription factor(NF-κB),rat cryptothermal protein(NLRP3)and other indexes of rats in each group were detected.The therapeutic effect of scrophularia-atractylodes and metformin on type 2 diabetic rats was preliminarily evaluated.The metabolic profiles of each group was further revealed by UPLC-MS/MC non-targeted metabolomics.The differential metabolites were screened and identified.Results:Compared with positive group,the treatment group had more obvious effects on improving the clinical symptoms and reducing postprandial hyperglycemia(P﹤0.05),and could significantly reduce the levels of NF-κB and NLRP3(P﹤0.01).A total of 36 differential metabolites were screened by metabonomics as serum potential biomarkers which were related to the mechanism of crophularia-atractylodes and metformin on diabetes mellitus.D-glutamine and D-glutamate were the related metabolic pathway which was involved in the treatment of diabetes mellitus with scrophularian-atractylodes and metformin.Conclusion:Scrophulariaatractylodes combined with metformin are effective on diabetes mellitus.It is speculated that the mechanism of scrophularian-atractylodes and metformin on diabetes mellitus may be to maintain the dynamic balance of"glutamine-glutamate"by regulating the expression of genes of D-glutamine and D-glutamate metabolic pathways,and to inhibit the activation of NF-κB/NLRP3 pathway,so as to improve the body's sensitivity to insulin,regulate the disorder of glucose and lipid metabolism,and reduce the reaction of inflammation.