黄芪黄酮干预脾虚水湿不化大鼠血浆代谢组学研究

采用高效液相色谱结合飞行时间质谱检测正常大鼠、脾虚水湿不化证大鼠及黄芪黄酮组分干预后大鼠血浆内源性代谢物变化,获取大鼠血浆代谢图谱,研究黄芪黄酮组分干预脾虚水湿不化证的作用机制.采用高脂低蛋白饮食加负重游泳力竭建立脾虚水湿不化证大鼠模型,选择Halo C18色谱柱,流动相为0.05％甲酸-水与0.05％甲酸-乙腈,梯度洗脱,利用液相色谱-串联质谱分析测定大鼠血浆样品.利用主成分分析法对造模前后大鼠血浆样本进行代谢轮廓分析,结合变量投影重要性及显著性分析等筛选对分组贡献最大的差异标志物及相关通路,阐明药物的作用机制.结果表明, 黄芪黄酮组代谢轮廓异于模型组,而接近于正常组,共鉴定出了包含甘油磷脂类、鞘酯类、氨基酸类等在内的11种潜在生物标志物,其代谢通路包括三羧酸循环、甘油磷脂代谢、脂肪酸代谢及氨基酸代谢等,主要涉及体内能量代谢和脂肪代谢.黄芪黄酮干预脾虚水湿不化证大鼠后, 宏观指标(如体重、自主活动)和微观指标(如代谢标志物、血脂)均明显回调,表明黄芪黄酮可能主要通过调节能量代谢、脂肪代谢等途径而发挥健脾利水作用.

Plasma Metabonomics Research of Astragalus Flavonoids Intervention on Rats of Dampness Stagnancy Due to Spleen Deficiency

A method of high performance liquid chromatography coupled with time-of-flight mass spectrometry was used to detect the endogenous metabolites changes in plasma of normal rats, rats of dampness stagnancy due to spleen deficiency and Astragalus flavone component intervention rats. Metabolism map of rat plasma was obtained and the mechanism of Astragalus flavonoids on dampness stagnancy due to spleen deficiency was studied. Rat model with dampness stagnancy due to spleen deficiency was established by high fat and low protein diet plus load swimming. Liquid chromatography tandem mass spectrometry was used for the analysis of rat plasma sample, and 0.05％ formic acid water with 0.05％ formic acid acetonitrile as the mobile phase was applied in gradient elution with Halo C18 chromatographic column. In this study, partial least squares discriminant analysis and variance analysis were used to screen the potential biomarkers, it was found that the metabolic profile of the Astragalus flavonoids was different from that of the model group, which was close to that of the normal group. A total of 11 potential biomarkers were identified, including glycerol phospholipids, sphingolipids, amino acids, and so on. The metabolic pathways of biomarkers including three tricarboxylic acid cycle, glycerol phospholipid metabolism, fatty acid metabolism, amino acid metabolism and so on, which mainly related to energy metabolism and fat metabolism in the body. Related indexes of rats with syndrome of spleen deficiency of water and dampness were significantly callback after Astragalus flavone intervention, including macro indicators such as body weight, independent activities and micro indicators such as metabolic markers, blood lipids and others. The result showed that Astragalus flavonoids played the role of strengthening the spleen and draining the water mainly through regulating the energy metabolism, fat metabolism and so on.