Mechanisms of Shengxian Quyu Decoction In Heart Failure Using an Untargeted Metabolomics and Network Pharmacology Approach In Rats

Summary

This study confirmed the therapeutic effect of Shengxian Quyu Decoction on heart failure through animal experiments. Ultra-performance liquid chromatography–mass spectrometry and network pharmacology analyses identified potential therapeutic targets and mechanisms.

Abstract

Shengxian Quyu decoction (SXQY) has been suggested as a potential therapeutic strategy for heart failure (HF), but its therapeutic mechanisms remain unclear. This study investigated the therapeutic effects and underlying mechanisms of SXQY in HF. A rat model of HF was induced by transverse aortic constriction (TAC) and treated with SXQY. Cardiac function was assessed by transthoracic echocardiography, and myocardial structure and fibrosis were evaluated using hematoxylin and eosin and Masson’s trichrome staining. Serum samples were analyzed by untargeted metabolomics using ultra-performance liquid chromatography–tandem mass spectrometry. Blood-entering components were mapped to targets, and intersecting HF-related targets were analyzed using protein–protein interaction (PPI) network analysis, followed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. SXQY significantly improved cardiac function in TAC rats, as evidenced by decreased left ventricular internal diameter in diastole and left ventricular posterior wall thickness in diastole and increased left ventricular ejection fraction, and alleviated myocardial hypertrophy, inflammation, and fibrosis. A total of 563 blood-entry compounds were identified (367 prototypes and 196 metabolites), with 5 key active compounds identified in the Traditional Chinese Medicine Systems Pharmacology database corresponding to 63 targets. Network analysis revealed 44 overlapping genes, with Formononetin, Timosaponin BII, and Sinensetin as core components, and PTGS2, PPARG, and HSP90AA1 as hub targets. PPI analysis further identified key genes including ESR1, PTGS2, PPARG, HSP90AA1, JUN, and 15 additional genes. KEGG analysis indicated that SXQY mainly acts via Ca²⁺, phosphatidylinositol 3-kinase–protein kinase B, cyclic adenosine monophosphate, and inflammation- and hormone-related pathways. In conclusion, SXQY exerts protective effects against HF by improving cardiac function and attenuating myocardial remodeling through multicomponent, multitarget, and multipathway mechanisms.

Introduction

Heart failure (HF) is a clinical syndrome characterized by impaired ventricular filling and ejection capacity due to cardiac structural or functional abnormalities, resulting in a constellation of clinical signs and symptoms¹. As the terminal stage of various cardiac diseases, HF significantly compromises patients’ quality of life and results in high mortality and frequent hospital readmissions, thereby imposing a substantial disease burden on both families and society^2,3. In clinical management, pharmacotherapy for HF forms the foundation and cornerstone of treatment, primarily c....

Protocol

All experimental procedures were examined and approved by the Animal Ethics and Welfare Committee of China–Japan Friendship Hospital (approval No. zryhyy21-23-01-09) and were conducted in accordance with the replacement, reduction, and refinement principles to minimize animal use and suffering. Investigators performing data acquisition and analysis (including echocardiography measurements and histological assessment) were blinded to group allocation.

Preparation of SXQY sample

SXQY is composed of the following herbal components: Astragalus membranaceus (Huangqi) (30 g), Codonop....

Results

Transthoracic Echocardiography

Echocardiographic analysis revealed pronounced cardiac remodeling and dysfunction in TAC rats (Figure 1). Compared with Sham controls, TAC significantly increased LVIDd and LVPWd, while LVEF was markedly decreased. Notably, SXQY administration partially reversed these pathological changes, as reflected by a significant reduction in LVIDd and LVPWd and a concomitant improvement in LVEF compared with the TAC group.

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Discussion

SXQY has been extensively used in clinical practice for over two decades, demonstrating efficacy in treating various cardiovascular conditions. Recently, its use has been extended to patients with heart failure with reduced ejection fraction and mildly reduced ejection fraction¹⁶. In the present study, a rat model was established by TAC surgery to mimic pathological changes of HF, and meanwhile some rats were treated with SXQY to examine its efficacy. Transthoracic echocardiography, HE staining, a.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
2-chloro-L-phenylalanine	Aladdin	C105993-1g	Internal standard; ≥98%
Acetonitrile (LC-MS grade)	Aladdin	A120771-1L	Extraction solvent
Acetic acid	Solarbio	G1340	Masson staining; pre-cooled to 4 °C
Autosampler vials	Agilent	5190-9590	For LC-MS/MS sample loading
Buprenorphine	Cerilliant	B-044S	Analgesic
C18 column (1.8 μm, 2.1 mm × 100 mm)	Waters	176001125	ACQUITY Premier HSS T3 column
Capillary tubes, sterile	Fisherbrand	22-362-566	Retro-orbital blood collection
Centrifuge (refrigerated)	KUBOTA	N61196-F000	For serum and sample processing
clusterProfiler package	Bioconductor	—	R package for KEGG enrichment analysis; v4.0.2
Curved forceps, sterile	HL-SQX	HL-JP	Aortic arch manipulation
Cytoscape software	NHGRI	NIH NHGRI U24 HG012107	Version 6.4.1
Dedicated imaging platform	Fujifilm VisualSonics	VisualSonics Vevo 1100 ultrasound imaging system	Animal positioning during echocardiography
Distilled water	N/A	N/A	Decoction preparation and vehicle control
Electric heating mantle	Midea	WK2102	Decoction boiling
Endotracheal tube	CWE Inc., USA	SAR-1000	Endotracheal intubation; The endotracheal tube used in this study was integrated into the ventilator configuration and was not a separate or standalone device
Ethanol (absolute)	Beijing Yili Fine Chemicals Co., Ltd.	64-17-5	Dehydration and sterilization
Formic acid	Aladdin	O930702	Mobile phase additive
Gauze	Solarbio	YA0720	Decoction filtration; 200–300 μm pore size
GeneCards database	GeneCards	—	Disease-related target retrieval; accessed January 2026
GNPS database	University of California San Diego	—	Spectral database; version 1.3.16, data archived in December 2025
GraphPad Prism software	GraphPad Software	—	Version 10.1.2; SCR_002798
Heating pad (thermostatic)	Physitemp	HP-4M	Maintained at 37 °C
Hematoxylin and Eosin (H&E) Staining Kit	Solarbio	G1120	Histological staining
High-frequency transducer	VisualSonics	Vevo F2	18–40 MHz
High-resolution mass spectrometer	Thermo Fisher Scientific	Q Exactive	Q Exactive HF-X
High-resolution small-animal ultrasound system	Fujifilm VisualSonics	VisualSonics Vevo 1100 ultrasound imaging system	Vevo 2100
HMDB database	Wishart Research Group / The Metabolomics Innovation Centre, University of Alberta	—	Spectral database; HMDB current version 5.0; release 5.0; January 2022
In-house reference standard library	METWARE	—	Composition of the in-house reference standard library: The library contains a total of 5,000 compounds and includes both MS1 (precursor) information and MS2 (fragmentation) spectral data. Availability statement: The library data format is .txt. Storage condition: The database is stored on the company's internal server. Access permissions: This self-built standard library is a proprietary in-house spectral library of the company. It is used solely for metabolite identification, result verification, and quality traceability within the company's controlled analytical workflows. The library is not publicly available for download. Clients may obtain annotation results, hit information, and necessary field descriptions generated based on this library.
Isoflurane	Sigma-Aldrich	792632	Inhalation anesthetic
Liquid chromatography system (UPLC)	SCIEX	ExionLC AD	ExionLC AD
MassBank database	MassBank consortium; Mass Spectrometry Society of Japan / MassBank Europe	—	Spectral database; version 2024.06; data archived in December 2025
Masson’s Trichrome Stain Kit	Solarbio	G1340	Histological staining
Membrane filter (0.22 μm)	Beyotime	FF362	Polyethersulfone (PES)
METLIN database	The Scripps Research Institute / Siuzdak Lab	RRID: SCR_010500	Spectral database; this database does not provide a fixed annual version number; data downloaded in 2023
Methanol (LC-MS grade)	Beijing Yili Fine Chemicals Co., Ltd.	67-56-1	Extraction solvent
Microcentrifuge tubes (1.5 mL)	Corning Life Sciences (Wujiang) Co., Ltd.	MCT-150-C	Sample processing
Microtome	Leica	DSC1	Tissue sectioning
Neutral resin	Solarbio	G8590	Slide mounting
Nitrogen evaporator/gas source	Organomation	N-EVAP	Sample drying
Ophthalmic ointment	Aladdin	08-03-8009	Eye protection during anesthesia
Paraffin	Solarbio	YA0010	Tissue embedding
Paraformaldehyde (4%)	Solarbio	P1110	Tissue fixation
Phosphate-buffered saline	DU YOU YOU	DYP1021	Tissue washing
Phosphomolybdic acid	Solarbio	G1340	Masson staining
Povidone–iodine	Solarbio	IP3800	Surgical disinfection
ProteoWizard software	ProteoWizard Foundation	—	Version 3.0.19254
R software	R Foundation	—	Version 4.2.2
Rotary evaporator	QINGDAO JINGCHENG INSTRUMENT CO., LTD	JC-2LZF	Decoction concentration under reduced pressure
Silk suture (2-0)	FULAN	NL30RH26A-76	Aortic constriction
Small-animal ventilator	CWE Inc., USA	SAR-1000	Mechanical ventilation
Sprague–Dawley rats (male, 8 weeks)	SPF Bio Co., Ltd.	—	RGD_70508; License No. SCXK(jing)2024-0001
STRING database	STRING Consortium	—	Protein interaction analysis; accessed January 2026
Support vector regression normalization package/script	MetNormalizer	—	Signal drift correction; v1.3.02
TCMSP database	TCMSP	—	Compound and target retrieval; accessed January 2026
UniProt database	UniProt Consortium	—	Gene symbol standardization
Ultrasound gel	MAGNAFLUX	25-912	Echocardiography
Ultrapure water	Laboratory-prepared	—	Mobile phase solvent
Vortex mixer	Yalin	VM-ST	Sample mixing
Water bath	Julabo	PURA22	Section floating at 45 °C
Weishengxin platform	Weishengxin	—	Target intersection analysis; accessed January 2026
XCMS software	R Foundation	—	Version 3.16.1
Xylene	Aladdin	X775162	Deparaffinization