基于GC-MS技术的大鼠糖尿病发生发展及银杏叶提取物干预的血浆代谢组学研究

Metabolomics analysis for the occurrence and development of diabetic rat and intervention of Ginkgo biloba leaves extract based on gas liquid chromatography

目的利用代谢组学技术探寻糖尿病发生发展及银杏叶提取物干预的潜在生物标志物及机制。方法采用腹腔注射链脲佐菌素构建SD大鼠糖尿病模型,随机分为正常对照组(NC)、正常银杏叶提取物处理组(CG)、糖尿病模型组(DM)、糖尿病模型银杏叶提取物处理组(GBE)。各组大鼠持续干预9周,收集第6-9周各组大鼠血浆,利用气相色谱-质谱联用技术(GC-MS)进行分析,对所得数据进行主成分分析(PCA)及偏最小二乘法判别分析(PLS-DA),以筛选潜在生物标志物。结果PCA得分图显示,NC组和CG组无法区分,而NC组和DM组、DM组和GBE组可分别聚类,表明DM大鼠血浆中代谢物发生了异常变化,而GBE可改善DM大鼠血浆代谢物的异常。综合PLS-DA分析、t检验和倍数变化分析结果,在DM组大鼠血浆中分别筛选并初步鉴定出13个发生持续性变化的代谢产物;GBE逆转了其中的10个代谢产物。结论碳水化合物、脂肪酸、氨基酸、磷脂、三羧酸循环、有机酸等代谢异常推动DM的发生发展,而GBE可通过影响碳水化合物、脂肪酸、氨基酸、磷脂、三羧酸循环等代谢异常发挥其延缓DM发生发展的作用。

Objective To explore the biomarkers and potential mechanism in the occurrence and development of diabetes mellitus(DM) and intervention of Ginkgo biloba leaves extract(GBE) using metabolomics.Methods Diabetic rat was induced by intra-peritoneal injection of streptozotocin.The rats were randomly divided into normal control group treated with free-drug solution medium(NC),normal control group treated with GBE(CG),DM model group treated with free-drug solution medium(DM) and DM model group treated with GBE(GBE),and respectively intervened with the relevant medium for 9 weeks.The rat plasma was collected from the sixth week to ninth week and analyzed with gas chromatography coupled with mass spectrometry(GC-MS).The GC-MS data was respectively processed with principal component analysis(PCA) and partial least square discriminant analysis(PLS-DA) to screen the differential metabolites.Results The PCA scores showed that NC group could not be separated from CG group while DM group and NC group, and GBE group and DM group could be well separated.Those results hinted that the plasma metabolites in DM rat were significantly changed when comparing with NC group, while GBE could obviously improve the variation of plasma metabolites in DM rat but had no significant influence on the plasma metabolites in NC rat.The results from PLS-DA,t-test and fold change showed that 13 plasma metabolites in DM rat with continuous variation were screened and preliminarily identified, while GBE reversed the variation of 10 metabolites.Conclusion The results of metabolic pathway analysis showed that the dysfunction of carbohydrate, fatty acid, phospholipids, amino acid, tricarboxylic acid cycle and organic acid metabolism promoted the occurrence and development of DM,while GBE could prevent the occurrence and development of DM by improving carbohydrate, fatty acid, phospholipids, amino acid and tricarboxylic acid cycle metabolism.