基于代谢组学探讨血府逐瘀汤对冠心病心血瘀阻证模型大鼠的疗效机制

Efficacy and mechanism of Xuefu Zhuyu Decoction on model rats of coronary heart disease with heart blood stasis syndrome based on metabolomics

研究血府逐瘀汤对冠心病心血瘀阻证模型大鼠心肌代谢产物的影响,探讨活血化瘀法疗效机制。SD大鼠随机分为假手术组、模型组、血府逐瘀汤组(14.04 g·kg^(-1))、曲美他嗪组(5.4 mg·kg^(-1)),假手术组只穿线不结扎,其余组通过冠状动脉左前降支结扎法制备冠心病心血瘀阻证模型,造模3 d后进行药物干预,干预14 d后取材。观察一般状态,检测心电图、心脏彩超指标,苏木精-伊红(hematoxylin-eosin,HE)染色、Masson染色观察组织病理形态学,酶联免疫吸附测定法(enzyme linked immunosorbent assay,ELISA)检测血清甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)含量,利用超高效液相色谱-质谱联用(ultra high performance liquid chromatography-quantitative exactive-mass spectrometry,UHPLC-QE-MS)技术筛选心肌组织差异代谢物,并进行代谢通路富集分析。结果表明,血府逐瘀汤可以显著改善模型大鼠的一般状态,降低心电图心率、ST段抬高幅度,升高左室射血分数(left ventricular ejection fraction,LVEF)和短轴缩短率(left ventricular fractional shortening,LVFS),降低左室舒张末内径(left ventricular intemal diameter in diastole,LVIDd)和左室收缩末内径(left ventricular intemal diameter in systole,LVIDs);HE染色与Masson染色显示,血府逐瘀汤可有效减轻模型大鼠的心肌组织结构紊乱、炎性细胞浸润与胶原纤维沉积;ELISA结果显示,血府逐瘀汤有效调节模型大鼠的血清TG、TC水平。各组心肌样本的代谢表型差异明显,共鉴定出血府逐瘀汤组回调的14种差异代谢物,涉及5条代谢通路,包括精氨酸和脯氨酸代谢,甘油磷脂代谢,氨酰基tRNA生物合成,醚脂代谢,丙氨酸、天冬氨酸和谷氨酸代谢。血府逐瘀汤改善冠心病心血瘀阻证模型大鼠的心功能、心肌结构损伤,其生物学机制涉及调节脂质代谢、胆碱代谢、氨基酸代谢、能量代谢、蛋白质合成等相关途径。

This study investigated the effects of Xuefu Zhuyu Decoction on myocardial metabolites in a rat model of coronary heart disease with heart blood stasis syndrome and explored the therapeutic mechanism of blood circulation-promoting and blood stasis-removing therapy.SD rats were randomly divided into a sham operation group,a model group,a Xuefu Zhuyu Decoction group(14.04 g·kg^(-1)),and a trimetazidine group(5.4 mg·kg^(-1)).The sham operation group underwent thread insertion without ligation,while the other groups underwent coronary artery left anterior descending branch ligation to induce a model of coronary heart disease with heart blood stasis syndrome.Three days after modeling,drug intervention was performed,and samples were taken after 14 days of intervention.General conditions were observed,and electrocardiogram and cardiac ultrasound indices were measured.Hematoxylin-eosin(HE) staining and Masson staining were used to observe tissue pathological morphology.The enzyme linked immunosorbent assay(ELISA) was used to measure the levels of triglyceride(TG) and total cholesterol(TC) in the serum.Ultra high performance liquid chromatography-quantitative exactive-mass spectrometry(UHPLC-QE-MS) technology was used to screen differential metabolites in myocardial tissue and conduct metabolic pathway enrichment analysis.The results showed that Xuefu Zhuyu Decoction significantly improved the general condition of the model rats,reduced heart rate and ST segment elevation in the electrocardiogram,increased left ventricular ejection fraction(LVEF) and left ventricular fractional shortening(LVFS),and decreased left ventricular internal diameter in diastole(LVIDd) and left ventricular internal diameter in systole(LVIDs).HE staining and Masson staining showed that Xuefu Zhuyu Decoction effectively alleviated myocardial tissue structural disorders,inflammatory cell infiltration,and collagen fiber deposition in the model rats.ELISA results showed that Xuefu Zhuyu Decoction effectively regulated serum TG and TC levels in the model rats.There were significant differences in the metabolic phenotypes of myocardial samples in each group.Fourteen differential metabolites were identified in the Xuefu Zhuyu Decoction group,involving five metabolic pathways,including arginine and proline metabolism,glycerophospholipid metabolism,aminoacyl-tRNA biosynthesis,ether lipid metabolism,and alanine,aspartate,and glutamate metabolism.Xuefu Zhuyu Decoction improved cardiac function and myocardial structural damage in the rat model of coronary heart disease with heart blood stasis syndrome,and its biological mechanism involved the regulation of lipid metabolism,choline metabolism,amino acid metabolism,energy metabolism,and protein synthesis pathways.