Study On The Mechanism Of Lingguizhugan Decoction In The Treatment Of Heart Failure Based On Network Pharmacology And Metabolomics

Lingguizhugan Decoction(LGZGD),documented in Jinguiyaolue written by Zhongjing Zhang,is composed of Fulin,Guizhi,Baizhu and Gancao.It is clinically used in the treatment of heart failure(HF)with remarkable curative effect.Due to the complex constituents of LGZGD,the mechanism of LGZGD in the treatment of HF has not been fully elucidated.The mechanism researches of traditional pharmacology were mostly point-to-point type,which was single component aimed at single target or pathway.It is difficult to expound thse mechanism of traditional Chinese medicine,which were multicomponent aimed at multiple targets.Metabolomics and network pharmacology are approaches of holistic perspective and are in line with the systematic view of traditional Chinese medicine.Therefore,they are particularly suitable for studying the mechanism of traditional Chinese medicine:Objective: To investigate the therapeutic mechanism of LGZGD in HF treatment based on an integration of the metabolomics and network pharmacy at the metabolic level and gene level.Methods: The constituents of LGZGD were recognized and identified by application of ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometer(UHPLC-Q-TOF-MS)technology.The chemical standard,retention time,and cracking rules of MS information were used to identify the constituents.Using online platform Swiss Target Prediction,the targets of constituents of LGZGD were predicted.The targets of HF were obtained from online database Dis Ge NET.The shared targets were intersection of targets of LGZGD and targets of HF.The network of constituents-shared targets and protein-protein interactions(PPI)of shared targets were constructed in Cytoscape.The core targets and their related costituents were obtained then.Enrichment analysis and pathway analysis were applied on Metascape,and the mechanism of LGZGD in treatment of HF were predicted.Metabolomics method based on UHPLC-Q-TOF-MS was used to study the mechanism of LGZGD in treatment of HF.First,non-target metabolomics method was used to analyze serum metabolic profile of normal mice after administration of LGZGD.The differential metabolies were screened out and pathways of them were analysis.Second,non-target metabolomics method was used to analyze serum metabolic profile of doxorubicin-induced HF mice after administration of LGZGD.The efficacy of LGZGD was evaluated by integration of ultrasonic echocardiography,histopathological staining and biochemical assays.The differential metabolies were screened out and pathways of the them were analyzed.In addition,core metabolites were screened out by network analysis to predict the core target of LGZGD.Finally,lipidomics method was used to analyze serum lipid metabolic profile of doxorubicin-induced HF mice after administration of LGZGD.The differential lipid metabolies were screened out and pathways of the them were analysis.Results: Altogether 78 constituents were identified accurately or putatively in LGZGD,194 targets of constituents were predicted and 814 targets of HF were obtained.46 shared targets of LGZGD and HF were gained after intersection.In the network of constituents-shared targets and PPI,it was showed that the key targets,such as AKT1、TNF、PTGS2(COX-2)、EGFR、PPARG、STAT3、ACE、AGTR1 and et al,were possibly regulated by variety of constituents of LGZGD,involving regulation of IL-17,TNF,arachidonic acid signaling pathway and lipolysis regulation of lipids and lipoproteins process and et al.83 differential metabolies were identified in normal mice non-targets metabolomics study.It showed that alpha-linolenic acid,sphingolipid and glycerophospholipid metabolic pathways were involved in LGZGD effect.54 differential metabolies were identified in non-targets metabolomics of LGZGD in treatment of HF study.It showed that arachidonic and glycerophospholipid metabolic pathways were involved in LGZGD effect.After visual network analysis,the potential targets of LGZGD in the treatment of HF were predicted,including PLA2,PLD,PLB,ALT,AST,TAT,phenylalanine-4-mono-oxygenase and peroxidase.90 differential lipid metabolies were identified in lipidomics of LGZGD in treatment of HF study,in which arachidonic and glycerophospholipid metabolic pathways were involved.It is suggested that LGZGD might inhibit the activation of LOX and COX pathways and regulate lipid metabolism.Conclusion: By integration of metabolomics and network pharmacology in this study,it is showed that glycerophospholipid and arachidonic acid metabolism were significant involved as the mechanism of LGZGD in the treatment of HF.By combination all parts of results,it revealed that PLA2-LOX/COX-2 pathway were regulated by varieties of constituents of LGZGD,targeting PLA2,COX-2 and LOX.This study provided a theoretical basis for the mechanism of LGZGD in the treatment of heart failure.