基于代谢组学探讨加参方对慢性心力衰竭模型大鼠的作用及机制

Effect and mechanism of Jiashen Prescription on chronic heart failure model rats based on metabonomics

关键词:  加参方  慢性心力衰竭  代谢组学

Key words:  Jiashen Prescription  chronic heart failure  metabonomics

摘要:

[目的] 在加参方治疗慢性心力衰竭（CHF）的药效基础上，利用代谢组学技术探究其抗心力衰竭的作用机制。[方法] 采用结扎大鼠左冠状动脉前降支造成心肌梗死后形成CHF模型，连续给药4周，通过超声检测心脏功能、苏木精-伊红（HE）染色检测心肌病理变化、马松（Masson）染色检测心肌纤维化、酶联免疫吸附剂测定（ELISA）检测心肌组织N-末端脑利钠肽原（NT-proBNP）水平，探究加参方对CHF的药效作用；采用超高效液相色谱-飞行时间质谱联用技术（UPLC-Q-TOF-MS）技术对大鼠血清和尿液中小分子代谢物进行非靶向代谢组学检测并进行多元统计分析，寻找疾病发生的代谢标志物，最后采用MetaboAnalyst代谢组学分析平台构建加参方治疗CHF的代谢通路。[结果] 加参方可显著升高CHF大鼠心脏左室射血分数（EF）和左室短轴缩短率（FS），减轻CHF大鼠心肌病理损伤和心肌纤维化，降低心肌胶原容积分数（CVF），降低CHF大鼠心肌组织NT-proBNP水平。CHF大鼠血清得到33个代谢标志物，主要为脂质和氨基酸及其衍生物，尿液得到38个代谢标志物，主要为氨基酸及其衍生物、长链脂肪酸和核苷类；与模型组比较，加参方组大鼠血清和尿液中分别有21个和29个代谢物有回调趋势。加参方主要通过甘油磷脂代谢、鞘脂代谢及丙氨酸、天冬氨酸和谷氨酸代谢通路干预血清代谢标志物，通过泛酸和CoA生物合成、组氨酸代谢及嘌呤代谢通路干预尿液代谢标志物。[结论] 加参方主要通过脂质代谢、氨基酸代谢、能量代谢和嘌呤代谢通路改善CHF。

Abstract:

[Objective] Based on the efficacy of Jiashen Prescription in the treatment of chronic heart failure(CHF),to explore the mechanism of its anti-heart failure effect by metabonomics. [Methods] The CHF model was formed after myocardial infarction caused by ligation of the anterior descending branch of the left coronary artery in rats,and was administered continuously for 4 weeks. Cardiac function was detected by ultrasound,myocardial pathological changes were detected by HE staining,myocardial fibrosis was detected by Masson staining,and the level of N-terminal brain natriuretic peptide(NT-proBNP) in myocardial tissue was detected by ELISA. To explore the pharmacodynamic effect of Jiashen Prescription on CHF;Ultra-high performance liquid chromatography-time-of-flight mass spectrometry (UPLC—Q—TOF-MS) was used for untargeted metabolomics detection of small molecule metabolites in serum and urine of rats and multivariate statistical analysis was performed to find metabolic markers for disease occurrence. Finally,MetaboAnalyst metabolic analysis platform was used to construct the metabolic pathway of Jiashen Prescription for CHF treatment. [Results] It significantly increased left ventricular ejection fraction(EF) and short axis shortening rate(FS) in CHF rats,alleviated myocardial pathological injury and fibrosis,decreased myocardial collagen volume fraction(CVF),and decreased the level of NT-proBNP in CHF rats. The 33 metabolic markers,mainly lipids and amino acids and their derivatives,were obtained in serum of CHF rats,and 38 metabolic markers,mainly amino acids and their derivatives,long-chain fatty acids and nucleosides,were obtained in urine of CHF rats. Compared with model group,21 metabolites and 29 metabolites in serum and urine of Jiashen Prescription group showed a callback trend,respectively. Jiashen Prescription mainly interferes with serum metabolic markers through glycerol phospholipid metabolism,sphingolipid metabolism, alanine,aspartic acid and glutamate metabolism pathways,and interferes with urine metabolic markers through pantothenic acid and CoA biosynthesis,histidine metabolism and purine metabolism pathways. [Conclusion] Jiashen Prescription can improve CHF mainly through lipid metabolism,amino acid metabolism,energy metabolism and purine metabolism pathways.