基于代谢组学探讨速效救心丸治疗稳定型心绞痛气滞血瘀证的作用机制

Study on the mechanism of Suxiao Jiuxin Pills in treating stable angina pectoris with qi stagnation and blood stasis based on metabonomics

目的鉴定速效救心丸干预稳定型心绞痛气滞血瘀证患者的差异代谢物、代谢通路,探讨速效救心丸相关治疗作用机制。方法选择2019年9月—2021年9月在东直门医院等接受义诊的患者和健康人,试验组纳入30例稳定型心绞痛气滞血瘀证患者,对照组纳入30名健康人。利用液相色谱-质谱技术对两组受试者进行血清代谢组学分析,采用多变量偏最小二乘回归分析模型的前2个主成分的VIP值,结合单变量分析差异倍数和P值筛选差异代谢物,基于京都基因与基因组百科全书数据库对差异代谢物进行代谢通路富集分析。结果共鉴定出试验组与对照组的差异代谢物18种,其中,4-羟基肉桂酰亚胺、胆绿素-Ⅸ-β、2-亚甲基谷氨酸等代谢物含量低于健康人,左旋肉碱、维生素BT、胍丁胺、柠康酸、鸟嘌呤、薄荷脑等代谢物含量高于健康人。对比试验组患者服用速效救心丸治疗前后的21种差异代谢物,治疗后细菌叶绿素a、植物鞘氨醇、4-氧视黄醇、神经酰胺、1-甲基鸟嘌呤等代谢物含量较治疗前下降,6-酮前列腺素E1、树脂毒素、前列腺素G2含量较治疗前升高。差异代谢物质的异常变化提示稳定型心绞痛患者体内可能存在氨基酸、脂质、视黄醇、嘌呤、卟啉和叶绿素、瞬时受体电位通道的炎症介质调节、血清素能突触等多种代谢途径的紊乱。结论稳定型心绞痛气滞血瘀证患者与健康人体内代谢物存在明显差异,速效救心丸具有双向调节体内血清代谢物的作用,从而发挥调节肠道菌群、维持肠道屏障、改善血脂水平、增加血管内皮功能的稳定、减轻组织细胞炎症反应、抗氧化等作用。

Objective To identify changes in metabolites in patients with stable angina pectoris(SAP)with qi stagnation and blood stasis,explore the differential metabolites and metabolic pathways associated with SAP,and determine the therapeutic mechanisms associated with Suxiao Jiuxin Pills.Methods Patients and healthy recruited from September 2019 to September 2021 in Dongzhimen Hospital and other departments.The experimental group included 30 SAP patients with qi stagnation and blood stasis,and the control group included 30 healthy individuals.Serum metabolomics analysis was performed on both groups using liquid chromatography/mass spectrometry(LC/MS),and the VIP values of the first two principal components of the multivariate partial least squares-discriminant analysis model combined with a univariate analysis of difference fold-change and P values were used to screen differentially expressed metabolites.Metabolic pathway enrichment analysis of differential metabolites was performed using the Kyoto Encyclopedia of Genes and Genomes database.Results A total of 18 significant differential metabolites were identified in the experimental and control groups;the levels of 4-hydroxycinnamimide,biliverdin-Ⅸ-β,and 2-methylene glutamic acid were lower in SAP patients than in healthy individuals,and the L-carnitine,vitamin BT,agmatine,citraconic acid,guanine,and menthol levels were higher.Twenty-one significant differential metabolites were identified in the experimental group before and after the administration of Suxiao Jiuxin Pills.The levels of bacterial chlorophyll-a,sphingosine,4-oxoretinol,ceramide,1-methylguanine,and other metabolites decreased after administration,while those of 6-ketoprostaglandin E1,resin toxin,and prostaglandin G2 increased.The abnormal changes in differential metabolites suggest that numerous metabolic pathways are disordered in SAP patients,including amino acids,lipids,retinol,purines,porphyrins,chlorophyll,inflammatory mediator regulation of transient receptor potential channels,and serotonergic synapses.Conclusion There are significant differences in metabolites between patients with SAP with qi stagnation and blood stasis and healthy people,and Suxiao Jiuxin Pills have positive and negative effects on regulating serum metabolites in vivo,thus regulating intestinal flora,maintaining the normal intestinal barrier,improving blood lipid levels,increasing the stability of vascular endothelial function,reducing tissue cell inflammation response,and inducing antioxidant effects.