气虚证模型大鼠尿液代谢组学特征

Urine Metabolomics Characteristics of Qi Deficiency Pattern Model Rats

目的运用代谢组学方法探讨气虚证的潜在生物学基础。方法将20只SD大鼠随机分为正常对照组和气虚证模型组各10只,气虚证模型组大鼠采用限制饮食+游泳力竭法制备气虚证大鼠模型,正常对照组大鼠自由饮水、正常进食。运用超高效液相色谱-质谱联用(UHPLC-QE-MS)的代谢组学技术对各组大鼠尿液代谢物进行检测。通过SIMCA(V15.0.2)软件对差异代谢物进行主成分分析(PCA)和正交-偏最小二乘判别分析(OPLS-DA),并绘制相对应的得分图。两组差异代谢产物进一步进行代谢路径拓扑分析,筛选富集性显著的代谢路径。结果与正常对照组比较,气虚证模型组的PCA与OPLS-DA得分图显示的代谢轮廓存在显著差异,气虚证模型组得到32种差异代谢产物,32种代谢产物共参与了9条代谢通路,其中嘧啶代谢、戊糖和葡萄糖醛酸酯的相互转化、磷酸戊糖途径为富集显著的3条代谢通路。结论气虚证大鼠的尿液代谢产物主要涉及氨基酸类、嘧啶类、多肽类、糖类等多个层面,其差异代谢产物可能是气虚证的重要标志物。

Objective To explore the potential biological basis of qi deficiency pattern through metabolomics. Methods Twenty SD rats were randomly divided into a normal control group and a qi-deficiency model group with 10 rats in each group. Restricted diet combined with exhausting swimming were used to develop a qi-deficiency model;rats in control group was free to drink water and take normal diet. Ultra-high performance liquid chromatography-mass spectrometry(UHPLC-QE-MS) was used to detect the urine metabolites of rats in each group. Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were performed on the differential metabolites by SIMCA(V15.0.2) software, and the corresponding score maps were drawn. The differential metabolites were further subjected to metabolic pathways for topological analysis, and significantly enriched pathways were identified. Results There were significant differences in scores of PCA and OPLS-DA of metabolic profiles between normal control group and model group. Thirty-two differential metabolites belonging to 9 metabolic pathways were identified in the model group, of which pyrimidine metabolism pathway, pentose glucuronate interconversion pathway, and pentose phosphate pathway were the mostly enriched ones. Conclusion The urinary metabolites of qi-deficiency rats mainly involve amino acids, pyrimidines, peptides, carbohydrates, and others;the differential metabolites may act as important markers of qi deficiency pattern.