Method Article

Exploring the Mechanism of San Jie Tong Mai Formula Against Atherosclerosis via Network Pharmacology and Proteomics

DOI:

10.3791/70080

⸱

April 17th, 2026

 ,  ,  ,  ,  , 
1Department of Cardiology, the Third Affiliated Clinical Hospital of Changchun University of Chinese Medicine, 2Department of Respiratory, The Affiliated Hospital of Changchun University of Chinese Medicine, 3The Affiliated Hospital of Changchun University of Chinese Medicine

In This Article

Summary

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This study demonstrates that San Jie Tong Mai Formula attenuates atherosclerosis in ApoE-/- mice by regulating Hsd11b1-mediated glucocorticoid metabolism, thereby improving dyslipidemia and vascular inflammation. Integrated network pharmacology, proteomics, and molecular docking analyses reveal a multi-component, multi-target synergistic mechanism underlying its anti-atherosclerotic effects.

Abstract

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This study aimed to elucidate the mechanism of San Jie Tong Mai Formula (SJTMF) against atherosclerosis (AS), a leading cause of cardiovascular morbidity. Using ApoE-/- mouse models, we demonstrated that SJTMF significantly inhibits AS plaque progression. Through an integrated network pharmacology and proteomics strategy, five core bioactive components were identified: beta-sitosterol, naringenin, luteolin, isorhamnetin, and 3beta-hydroxy-24-methylene-8-lanostene-21-oic acid. Concurrently, proteomics revealed 129 AS-related proteins that were differentially expressed. Molecular docking confirmed high-affinity binding interactions between these components and the key target Hsd11b1, with their binding energies all below -5 kcal/mol. Mechanistic investigations further revealed that SJTMF may regulate Hsd11b1-mediated glucocorticoid metabolism. This regulation contributes to significant amelioration of both dyslipidemia and vascular inflammation, thereby suppressing AS development. Collectively, this work demonstrates, for the first time, the innovative mechanism by which a traditional Chinese medicine formula exerts anti-AS effects through multi-component synergistic regulation of the Hsd11b1 target, offering new insights for therapeutic intervention.

Introduction

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Atherosclerosis (AS), a chronic inflammatory disease affecting systemic vasculature, constitutes the primary pathological basis for cardiovascular and cerebrovascular events1. Its pathogenesis involves complex interactions among genetic, environmental, psychosocial, and multifactorial pathological factors, exhibiting multistage and polygenic characteristics. Although current therapeutic strategies-particularly lipid-lowering agents such as statins and Proprotein Convertase Subtilisin/Kexin Type 9(PCSK9) inhibitors-effectively reduce low-density lipoprotein cholesterol (LDL-C) levels, they do not fully address the residual inflammatory risk that....

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Protocol

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All animal procedures were conducted with approval from the Experimental Animals Committee of Changchun University of Chinese Medicine (Ethics Approval No. 2023521). Specific pathogen-free (SPF) male C57BL/6J and ApoE⁻/⁻ mice (5 weeks old, 18-22 g) were included in the study (Laboratory Animal Use License Nos. SCXK (JI) 2023-002 and SCXK (SU) 2021-0013). Information regarding reagents and instrumentation is provided in the Table of Materials.

1. Network pharmacology

  1. Prediction of Active Components and Targets of SJTMF
    1. Screen the active components of Salvia ....

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Results

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SJTMF suppresses atherosclerotic plaque formation in ApoE-/- mice
To investigate the inhibitory effect of SJTMF on AS, ApoE-/- mouse model was used to monitor body weight changes and observe histopathological alterations in the aorta. Compared with the Model group, SJTMF intervention ameliorated body weight gain in HFD-fed ApoE-/- mice (Figure 1A). Specifically, different doses of SJTMF showed a trend tow.......

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Discussion

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Although previous studies have demonstrated that SJTMF exerts anti-inflammatory effects and induces macrophage polarization in ApoE⁻/⁻ mouse models6,7, the specific molecular mechanisms underlying its anti-atherosclerotic activity remain unclear. The "same treatment for different diseases" principle in traditional Chinese medicine stems from the synergistic effects of multi-component, multi-target, and multi-pathway regulation. Netw.......

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Disclosures

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The authors have no conflicts of interest to declare.

Acknowledgements

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This study was supported by the Natural Science Foundation of Jilin Province (Nos. YDZJ202401644ZYTS).

....

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
2-Chloroacetamide (CAA)Sigma-AldrichC0627-100G
4% Paraformaldehyde Fix SolutionSinopharm Chemical Reagent Co., Ltd.80096618
0.5–10 µL Micropipetteeppendorf4924000029
10 – 100 µL Micropipetteeppendorf4924000053
20 – 200 µL Micropipetteeppendorf4924000061
100 – 1,000 µL Micropipetteeppendorf4924000088
1000 µL Pipette TipsNEST Life Science Co., Ltd.303216
10 µL Pipette TipsNEST Life Science Co., Ltd.301016
20 µL Pipette TipsNEST Life Science Co., Ltd.302106
15 mL Centrifuge TubeNEST Life Science Co., Ltd.601052
1.5 mL Centrifuge TubeNEST Life Science Co., Ltd.615601
A high - throughput experiment - and reference - guided database of traditional Chinese medicine(HERB)/http://herb.ac.cn/v2
Acetonitrile (LC-MS)Thermo Fisher ScientificA955-4
Acetonyl acetoneSinopharm Chemical Reagent Co., Ltd.10000418
Ammonia WaterSinopharm Chemical Reagent Co., Ltd.10011028
AutoclaveShanghai Sanshen Medical Devices Co., Ltd.YM75
Acclaim PepMap (75µm*2cm, C18, 3µm, 100Å)Thermo164535
AURORA ULTIMATE (75µm*25cm, 1.6µm, 100Å)AURORAAUR3-25075C18-CSI
BCA Protein Assay KitSangon Biotech (Shanghai) Co., Ltd.C503021-0500
Benchtop High - Speed Refrigerated CentrifugeHunan Kecheng Instrument Equipment Co., Ltd.H1 - 16KR
Bromophenol BlueSinopharm Chemical Reagent Co., Ltd.71008060
Block HeaterHangzhou Miu InstrumentsDKT-100
ChloroformSinopharm Chemical Reagent Co., Ltd.10006818
Chemiluminescence Imaging SystemHangzhou Shenhua Technology Co., Ltd.SH - 523
Clean BenchSuzhou Jinghua Equipment Co., Ltd.SW-CJ-2D
Cover GlassJiangsu Shitai Experimental Equipment Co., Ltd.10212450C
Cytoscape v3.9.1//
DIA-NN (V1.8.1)The Francis Crick Institutegithub.com/vdemichev/DiaNN
Digital Display Constant Temperature Water BathChangzhou Jinnan Instrument Manufacturing Co., Ltd.HH - W600
Destaining ShakerWuhan Lingsi Biotechnology Co., Ltd.TSY-B
Dewatering machineWuhan Junjie Electronics Co., Ltd.JT-12J
EosinWuhan Lingsi Biotechnology Co., Ltd.E8090
Embedding ParaffinSinopharm Chemical Reagent Co., Ltd.69019361
Ethanol AbsoluteSinopharm Chemical Reagent Co., Ltd.10009218
Embedding MachineWuhan Junjie Electronics Co., Ltd.JB-P5
GraphPad Prism/8.0.2
Glacial Acetic AcidSinopharm Chemical Reagent Co., Ltd.10000218
GeneCards/https://www.genecards.org/
GlycerolSinopharm Chemical Reagent Co., Ltd.10010618
GlycineBiofroxx1275GR500
Gene Ontology (GO)/geneontology.org
HematoxylinWuhan Lingsi Biotechnology Co., Ltd.G1140
Horizontal ShakerHaimen Qilinbeier Instrument Manufacturing Co., Ltd., JiangsuTS - 1
Hydrophobic Barrier PenServicebioWG1066-1
HCIXinyang Chemical Reagent FactoryGB622-89
High-performance liquid chromatographyAgilent1200
IsopropanoSinopharm Chemical Reagent Co., Ltd.80109218
Image-Pro PlusMedia CyberneticsVersion 6.0
Imaging SystemNikonDS-Fi3
IRIS-Fine Micro Dissecting ScissorsRWD Life Science Co., Ltd.S12004-09
Kyoto Encyclopedia of Genes and Genomes (KEGG)/www.genome.jp/kegg/
Microscope SlideNantong Mevid Life Science Co., Ltd.PC2-301
Microwave OvenGalanz Microwave Oven Electrical Appliances Co., Ltd.P70D20TL-P4
Mini CentrifugeDragon LabD1008E
Neutral ResinWuhan Lingsi Biotechnology Co., Ltd.G8590
NaClSinopharm Chemical Reagent Co., Ltd.10019318
NCBI/www.ncbi.nlm.nih.gov
Non-contact ultrasonic cell pulverizerSCIENTZ08-III
n-ButanolSinopharm Chemical Reagent Co., Ltd.100052190
OvenShanghai Huitai Instrument Manufacturing Co., Ltd.DHG-9140A
PubChem/https://pubchem.ncbi.nlm.nih.gov/
PBS Phosphate Buffer PowderWuhan Lingsi Biotechnology Co., Ltd.P1010
Pathological MicrotomeLeica Instruments Co., Ltd.(Shanghai)RM2016
Portable CentrifugeWuhan Lingsi Biotechnology Co., Ltd.D1008
R 4.5.1//
RIPA Lysis Buffer (1.5 ML)MeilunbioMA0151
Sodium dodecyl sulfate (SDS)Sigma-AldrichL3771-500G
SpeedVacGemmyCV200
SPSS softwareIBM SPSS statisticsversion 22
Suture Tying ForcepsRWD Life Science Co., Ltd.FC11001R-11
Stereo Dissecting MicroscopeRWD Life Science Co., Ltd.77001S
Spin Columns RMCWBIOCW0597S
timsTOF Pro mass spectrometerBruker DaltonicstimsTOF Pro
Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP)/https://tcmspw.com/tcmsp.php
Thermo mixerHangzhou Miu InstrumentsMTC-100
Trifluoroacetic acid (TFA)Sigma-Aldrich299537-500G
Tris(2-carboxyethyl)phosphine hydrochloride (TCEP)Sigma-AldrichC4706-10G
TrypsinSigma-AldrichT575-31N-10
Tissue HomogenizerHangzhou Miu InstrumentsMT-30K
Tris(hydroxymethyl)aminomethane (Tris base)Sigma-AldrichT1503-1KG
UltiMate 3000 RSLCnano systemThermo ScientificUltiMate 3000 RSLCnano
UniProt (2023-01-03)The Francis Crick Institutegithub.com/vdemichev/DiaNN
UreaSigma-Aldrich51456-500G
Venny2.1/https://www.bioinformatics.com.cn/static/others/jvenn/example.html
Vortex MixerKylin-BellVORTEX-5
Vannas Spring-Loaded ScissorsRWD Life Science Co., Ltd.S11001-08
Water (LC-MS)Thermo Fisher ScientificW6-4
XyleneSinopharm Chemical Reagent Co., Ltd.10023418

References

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  1. Mensah, G. A., Roth, G. A., Fuster, V. The global burden of cardiovascular diseases and risk factors: 2020 and beyond. J Am Coll Cardiol. 74 (20), 2529-2532 (2019).
  2. Sampson, U. K., Fazio, S., Linton, M. F.

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Tags

Atherosclerosis MechanismSan Jie Tong MaiNetwork PharmacologyProteomics AnalysisApoE Knockout MiceBioactive ComponentsMolecular DockingGlucocorticoid MetabolismVascular InflammationHsd11b1 Target

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