基于非靶向代谢组学探讨麝香保心丸治疗冠心病心绞痛作用机制研究

Exploring the mechanism of action of Shexiang Baoxin Pills in treating angina pectoris based on untargeted metabolomics

关键词:  麝香保心丸  冠心病心绞痛  代谢组学  代谢通路

Key words:  Shexiang Baoxin Pill  angina pectoris of coronary heart disease  metabolomics  metabolic pathway

摘要:

目的 采用代谢组学方法探索麝香保心丸治疗冠心病心绞痛的作用机制，发掘与鉴定其潜在的生物标志物。方法 纳入冠心病心绞痛患者60例，将患者分为麝香保心丸组（麝香保心丸联合西医常规治疗）和西医常规治疗组（单独使用西医常规治疗），每组各30例，同时纳入正常对照组30例。采用超高效液相色谱-串联质谱（UPLC-MS/MS）技术对冠心病心绞痛患者治疗前后血清样本进行代谢组学检测，采用主成分分析（PCA）及正交偏最小二乘法判别分析（OPLS-DA）进行多元统计分析，结合MetaboAnalyst数据库寻找各组差异代谢物与相关代谢通路。采用ROC曲线评估上述筛选出的代谢物对冠心病心绞痛的诊断价值。结果 与正常对照组相比，冠心病心绞痛患者在PCA及OPLS-DA模型散点图上呈现明显的分类聚集，说明其代谢模式有较大的差异。冠心病心绞痛患者血清中的α-卡茄碱、华蟾毒精、全反式六杂环二磷酸酯、灵芝酸α等12种代谢物较正常对照组发生改变。ROC曲线证明上述12个潜在的差异代谢物对于冠心病心绞痛的诊断具有良好的效能。其中，麝香保心丸组有11种与上述冠心病心绞痛疾病相关的差异代谢物发生回调，分别为α-卡茄碱、华蟾毒精、SM[d20∶1/20∶5（6E，8Z，11Z，14Z，17Z）-OH（5）]、TG[14∶0/20∶0/14∶1（9Z）]、灵芝酸α、丙酰辅酶A、全反式六杂环二磷酸酯、棕榈酰葡糖苷酸、羟基鞘磷脂C24∶1、LacCer（d18∶1/20∶0）、DG（14∶0/0∶0/14∶1n5），涉及包括鞘脂代谢、乙醛酸和二羧酸代谢、丙酸代谢、β-丙氨酸代谢、缬氨酸、亮氨酸和异亮氨酸代谢5条代谢通路。结论 麝香保心丸可能通过调控鞘脂代谢、乙醛酸和二羧酸代谢、丙酸代谢、β-丙氨酸代谢、缬氨酸、亮氨酸和异亮氨酸代谢发挥对冠心病心绞痛的治疗作用。

Abstract:

Objective To explore the mechanism of action of Shexiang Baoxin Pills(SBP) in the treatment of angina pectoris, and to identify and characterize potential biomarkers associated with its therapeutic effects.Methods A total of 60 patients with angina pectoris were enrolled. Patients were divided into two groups: the SBP group(SBP combined with conventional Western medicine treatment) and the Western medicine-only group(treated with conventional Western medicine alone), with 30 patients in each group. Additionally, 30 healthy individuals were included as the control group. Serum samples from the patients before and after treatment were analyzed using ultra-performance liquid chromatography-mass spectrometry(UPLC-MS/MS) for metabolomic profiling. Multivariate statistical analyses, including principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA), were conducted. The MetaboAnalyst database was used to identify differentially expressed metabolites and related metabolic pathways. To further assess the diagnostic significance of the selected metabolites for angina pectoris, receiver operating characteristic(ROC) curves were used.Results Compared with the healthy control group, angina pectoris patients exhibited significant clustering on the PCA score plot, indicating substantial differences in their metabolic profiles. A total of 12 metabolites, including α-carboxylic acid, bufotenin, all-trans hexahydro-2H-pyrrolo[3, 4-b]quinolin-2-one, and ganoderic acid α, were found to be altered in the serum of angina pectoris patients. The ROC curve analysis showed that these 12 potential biomarkers exhibited good diagnostic performance for angina pectoris. In the SBP group, 11 out of these 12 differentially expressed metabolites showed reversal, including α-carboxylic acid, bufotenin, all-trans hexahydro-2H-pyrrolo[3, 4-b]quinolin-2-one, ganoderic acid α, propionyl-CoA, SM [d20∶1/20∶5(6E, 8Z, 11Z, 14Z, 17Z)-OH(5)], palmitoyl glucuronide, TG [14∶0/20∶0/14∶1(9Z)], hydroxysphingolipid C24∶1, LacCer(d18∶1/20∶0), and DG(14∶0/0∶0/14∶1n5). These metabolites were associated with five major metabolic pathways: sphingolipid metabolism, aldehyde acid and dicarboxylic acid metabolism, propionate metabolism, β-alanine metabolism, and valine, leucine, and isoleucine metabolism.Conclusion SBP may exert its therapeutic effects in angina pectoris by regulating sphingolipid metabolism, aldehyde acid and dicarboxylic acid metabolism, propionate metabolism, β-alanine metabolism, and valine, leucine, and isoleucine metabolism.