基于代谢组学技术的冠心病血瘀证不同进展阶段大鼠潜在生物标志物及代谢通路研究

目的通过超高效液相色谱-飞行时间质谱(UPLC-TOF/MS)的非靶向代谢组学方法探索冠心病血瘀证形成过程中大鼠心肌组织代谢特征,寻找潜在的生物标志物,为基于代谢组学的心肌保护相关治疗措施寻找有意义的靶点。方法取27只SD大鼠作为模型组制备冠心病血瘀证模型,另设空白组8只。模型组大鼠采用高脂饲料喂养+维生素D3灌胃+皮下注射异丙肾上腺素方法分3个阶段进行冠心病血瘀证模型的逐步制备。通过UPLC-TOF/MS技术获取大鼠心肌组织内源性小分子代谢物,利用SIMCA 14.1软件和MetaboAnalyst5.0软件进行组间差异性代谢物筛选及相关代谢通路分析,使用SPSS21.0对组间差异性代谢物进行定量分析,受试者工作特征(ROC)曲线评估特征代谢物临床诊断效能。结果筛选出磷脂酰胆碱、磷脂酰乙醇胺、1-酰基-sn-甘油-3-磷酸胆碱、L-苯丙氨酸、牛磺酸共5种差异最为显著的潜在生物标志物,涉及甘油磷脂代谢,苯丙氨酸、酪氨酸和色氨酸的生物合成,苯丙氨酸代谢,牛磺酸和亚牛磺酸代谢4条主要差异代谢通路。ROC曲线分析显示,特征代谢物曲线下面积均大于0.7。结论冠心病血瘀证不同进展阶段大鼠心肌组织...

Study on Potential Biomarkers and Metabolic Pathways of Rats in Different Progressive Stages of Coronary Heart Disease with Blood Stasis Syndrome Based on Metabonomics Technology

Objective To explore the metabolic characteristics and potential biomarkers of myocardial tissue during the formation of blood stasis syndrome in coronary heart disease through the non-targeted metabolomics method of ultra performance liquid chromatography-time of flight mass spectrometry(UPLC-TOF/MS);To look for potential biomarkers;To find meaningful targets for clinical myocardial protection-related treatment measures based on metabonomics.Methods Twenty-seven SD rats were set as model group to prepare rat models of coronary heart disease with blood stasis syndrome,with 8 rats as blank group.The rats in the model group were fed with high-fat diet+vitamin D3 gavage+subcutaneous injection of isoproterenol in three stages to gradually replicate the model of coronary heart disease blood stasis syndrome.The endogenous small molecule metabolites of rat myocardial tissues were obtained by UPLC-TOF/MS technology.SIMCA 14.1 software and MetaboAnalyst 5.0 software were used to screen different metabolites between groups and analyze related metabolic pathways.Differential metabolites between groups were quantitatively analyzed by SPSS 21.0 and the receiver operating characteristic(ROC)curve was used to evaluate the clinical diagnostic efficacy of characteristic metabolites.Results Phosphatidylcholine,phosphatidylethanolamine,1-acyl-sn-glycero-3-phosphocholine,L-phenylalanine,and taurine were screened for the five kinds of most significant difference potential biomarkers,involving four main different metabolic pathways:glycerophospholipid metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,phenylalanine metabolism,taurine and hypotaurine metabolism.ROC curve analysis showed that the areas under the curve of characteristic metabolites were all greater than 0.7.Conclusion Metabonomics of rat myocardial tissue changes in different stages of coronary heart disease with blood stasis syndrome.Its pathological mechanism mainly involves lipid metabolism and amino acid metabolism,and characteristic metabolites have diagnostic value.