基于UPLC/QTOF-MS技术的丹蛭降糖胶囊防治2型糖尿病血管病变大鼠的血清代谢组学研究

目的: 本实验运用UPLC/QTOF-MS技术对丹蛭降糖胶囊防治2型糖尿病血管病变大鼠进行血清的代谢组学研究,探求该中药制剂防治糖尿病血管病变可能的物质基础。方法: SD大鼠随机分成正常组、模型组和丹蛭降糖胶囊组,高脂饲料喂养4周后,腹腔注射小剂量链脲佐菌素(STZ35mg·kg^-1)复制2型糖尿病大鼠模型。造模成功后,灌胃给予丹蛭降糖胶囊(1.26g·kg^-1·d^-1)连续4周。给药4周后,处死大鼠,腹主动脉取血,去胸主动脉HE染色做病理切片。采用UPLC/QTOF-MS技术结合主成分分析(PCA)和偏最小二乘判别分析(PLS-DA)分析其代谢谱变化,鉴定差异化合物,分析相关代谢通路。结果: 病理切片发现模型组大鼠血管内皮细胞较正常组大鼠的有明显损坏,丹蛭降糖胶囊组大鼠的血管内皮细胞受损较轻。PCA和PLS-DA分析表明各组大鼠之间代谢谱存在明显差异,可实现分离。研究在正、负离子模式下分别筛选出11个和4个差异性标志物。代谢通路分析结果表明,差异性标志物分别于氨基酸代谢、胆汁酸和脂代谢以及氧化应激状态相关。结论: 丹蛭降糖胶囊可以保护血管内皮细胞,防治2型糖尿病血管病变,其作用机制可能在于调节氨基酸代谢、胆汁酸和脂代谢以及氧化应激状态相关。

Serum metabolomics of Danzhi Jiangtang Capsules against type 2 diabetic rats with vascular complications based on UPLC/QTOF-MS

OBJECTIVE To employ a serum metabolomics method based on UPLC/QTOF-MS to study possible metabolism and metabolic profile caused by Danzhi Jiangtang Capsules (DJC) against type 2 diabetic rats.METHODS All SD rats were randomly divided into normal group, type 2 diabetes model group and DJC group. Rats in type 2 diabetes model group and DJC group were fed with high-fat diet for four weeks after intraperitoneal injection of a small dose of streptozotocin (35 mg·kg^-1) to copy type 2 diabetic models. Then animal models were gavaged with DJC (1 260 g·kg^-1·d^-1) for 4 weeks, weekly monitoring blood glucose. After treatment, rat urine was collected, then all rats were sacrificed, abdominal aortic blood was used to test insulin and glycated hemoglobin in plasma. UPLC/QTOF-MS with principal component analysis (PCA) was employed to analyze metabolic profile changes, Metabolites were identified by using databases, and metabolism pathways were analyzed.RESULTS DJC could lower blood sugar, increase plasma insulin levels. Eleven potential metabolites in the positive ion mode and 4 potential metabolites in the negative ion mode were screened out, respectively. Potential metabolites were related to metabolism of amino acids, bile acids and lipids, and oxidation-antioxidant balance.CONCLUSION This study shows that DJC can lower blood glucose in diabetic rats and increase insulin levels possibly by regulating metabolism of amino acids, bile acids and lipids, and oxidation-antioxidant balance.