Mechanism of Ershen Zhenwu Decoction in ameliorating chronic heart failure via JNK/MAPK-regulated apoptosis: insights from network pharmacology and experimental validation

BackgroundChronic heart failure (CHF) is a complex cardiovascular disease caused by different pathological mechanisms. Modern medicine has made advancements in CHF treatment; however, there are still many challenges. Ershen Zhenwu Decoction (ESZWD) is a Xin'an medicine that has been clinically...

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Main Authors: Yulong Liu, Xinyue Wang, Maomao Zhang, Dan Cheng, Zhenpeng Zhu, Lan Ge, Xiaoyu Cheng
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Cardiovascular Medicine
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Online Access:https://www.frontiersin.org/articles/10.3389/fcvm.2025.1561963/full
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author Yulong Liu
Yulong Liu
Xinyue Wang
Xinyue Wang
Maomao Zhang
Maomao Zhang
Dan Cheng
Dan Cheng
Zhenpeng Zhu
Lan Ge
Xiaoyu Cheng
author_facet Yulong Liu
Yulong Liu
Xinyue Wang
Xinyue Wang
Maomao Zhang
Maomao Zhang
Dan Cheng
Dan Cheng
Zhenpeng Zhu
Lan Ge
Xiaoyu Cheng
author_sort Yulong Liu
collection DOAJ
description BackgroundChronic heart failure (CHF) is a complex cardiovascular disease caused by different pathological mechanisms. Modern medicine has made advancements in CHF treatment; however, there are still many challenges. Ershen Zhenwu Decoction (ESZWD) is a Xin'an medicine that has been clinically applied for years and had good efficacy against CHF; however, its underlying mechanisms remain undetermined. Therefore, this study aims to investigate the primary molecular mechanisms of ESZWD in CHF treatment and elucidate its multi-target and multi-level mode of action.ObjectiveThe aim of this study was to investigate the main molecular mechanisms of ESZWD for the treatment of CHF and to elucidate its multi-target and multi-level mode of action.MethodsThis study employed a network pharmacology approach to analyze the main ESZWD components and core targets. Furthermore, primary CHF targets were predicted to develop a protein–protein interaction (PPI) network and perform Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Moreover, molecular docking was carried out to validate the binding between active ingredients and key targets. For in vitro studies, myocardial cell injury models were employed, and immunofluorescence, RT-qPCR, Western blot, and flow cytometry were carried out to validate the critical targets of relevant signaling pathways and the specific ESZWD regulatory mechanisms.ResultsNetwork pharmacology identified 437 targets for 34 major ESZWD components. Of these, 216 drug–disease intersection targets were identified. The PPI network analysis identified the following core targets: STAT3, HSP90AA1, MAPK8, NFKB1, HIF1A, MMP9, PTGS2, BCL2L1, TLR4, and ESR1. GO analysis revealed that these targets were associated with exogenous stimuli responses, phosphorylation regulation, inflammatory response, and protein tyrosine kinase activity. Furthermore, KEGG analysis showed that ESZWD predominantly impacts cancer, inflammatory response, and apoptosis pathways, with c-Jun N-terminal kinase/mitogen-activated protein kinase (JNK/MAPK)-regulated apoptosis being a key pathway. In vitro analyses revealed that ESZWD effectively inhibited JNK activation, modulated MAPK signaling, downregulated pro-apoptotic gene expression, and significantly reduced cardiomyocyte apoptosis rates, thus validating the network pharmacology findings.ConclusionOur study shows that paeoniflorin, acetylaconitine, and cryptotanshinone bind to key proteins in the JNK/MAPK apoptosis pathway. In vitro validation confirms drug serum from ESZWD regulates this pathway, supporting its therapeutic potential for CHF.
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spelling doaj-art-b74f869efed44fae9cf6e7ec950d23872025-08-20T02:18:39ZengFrontiers Media S.A.Frontiers in Cardiovascular Medicine2297-055X2025-04-011210.3389/fcvm.2025.15619631561963Mechanism of Ershen Zhenwu Decoction in ameliorating chronic heart failure via JNK/MAPK-regulated apoptosis: insights from network pharmacology and experimental validationYulong Liu0Yulong Liu1Xinyue Wang2Xinyue Wang3Maomao Zhang4Maomao Zhang5Dan Cheng6Dan Cheng7Zhenpeng Zhu8Lan Ge9Xiaoyu Cheng10First Affiliated Hospital of Anhui University of Chinese Medicine, Anhui University of Chinese Medicine, Hefei, ChinaXin'an Key Laboratory of Medical Science, Anhui University of Chinese Medicine, Hefei, ChinaFirst Affiliated Hospital of Anhui University of Chinese Medicine, Anhui University of Chinese Medicine, Hefei, ChinaXin'an Key Laboratory of Medical Science, Anhui University of Chinese Medicine, Hefei, ChinaFirst Affiliated Hospital of Anhui University of Chinese Medicine, Anhui University of Chinese Medicine, Hefei, ChinaXin'an Key Laboratory of Medical Science, Anhui University of Chinese Medicine, Hefei, ChinaFirst Affiliated Hospital of Anhui University of Chinese Medicine, Anhui University of Chinese Medicine, Hefei, ChinaXin'an Key Laboratory of Medical Science, Anhui University of Chinese Medicine, Hefei, ChinaFirst Affiliated Hospital of Anhui University of Chinese Medicine, Anhui University of Chinese Medicine, Hefei, ChinaFirst Affiliated Hospital of Anhui University of Chinese Medicine, Anhui University of Chinese Medicine, Hefei, ChinaFirst Affiliated Hospital of Anhui University of Chinese Medicine, Anhui University of Chinese Medicine, Hefei, ChinaBackgroundChronic heart failure (CHF) is a complex cardiovascular disease caused by different pathological mechanisms. Modern medicine has made advancements in CHF treatment; however, there are still many challenges. Ershen Zhenwu Decoction (ESZWD) is a Xin'an medicine that has been clinically applied for years and had good efficacy against CHF; however, its underlying mechanisms remain undetermined. Therefore, this study aims to investigate the primary molecular mechanisms of ESZWD in CHF treatment and elucidate its multi-target and multi-level mode of action.ObjectiveThe aim of this study was to investigate the main molecular mechanisms of ESZWD for the treatment of CHF and to elucidate its multi-target and multi-level mode of action.MethodsThis study employed a network pharmacology approach to analyze the main ESZWD components and core targets. Furthermore, primary CHF targets were predicted to develop a protein–protein interaction (PPI) network and perform Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Moreover, molecular docking was carried out to validate the binding between active ingredients and key targets. For in vitro studies, myocardial cell injury models were employed, and immunofluorescence, RT-qPCR, Western blot, and flow cytometry were carried out to validate the critical targets of relevant signaling pathways and the specific ESZWD regulatory mechanisms.ResultsNetwork pharmacology identified 437 targets for 34 major ESZWD components. Of these, 216 drug–disease intersection targets were identified. The PPI network analysis identified the following core targets: STAT3, HSP90AA1, MAPK8, NFKB1, HIF1A, MMP9, PTGS2, BCL2L1, TLR4, and ESR1. GO analysis revealed that these targets were associated with exogenous stimuli responses, phosphorylation regulation, inflammatory response, and protein tyrosine kinase activity. Furthermore, KEGG analysis showed that ESZWD predominantly impacts cancer, inflammatory response, and apoptosis pathways, with c-Jun N-terminal kinase/mitogen-activated protein kinase (JNK/MAPK)-regulated apoptosis being a key pathway. In vitro analyses revealed that ESZWD effectively inhibited JNK activation, modulated MAPK signaling, downregulated pro-apoptotic gene expression, and significantly reduced cardiomyocyte apoptosis rates, thus validating the network pharmacology findings.ConclusionOur study shows that paeoniflorin, acetylaconitine, and cryptotanshinone bind to key proteins in the JNK/MAPK apoptosis pathway. In vitro validation confirms drug serum from ESZWD regulates this pathway, supporting its therapeutic potential for CHF.https://www.frontiersin.org/articles/10.3389/fcvm.2025.1561963/fullErshen Zhenwu decoctionnetwork pharmacologymolecular dockingJNK/MAPKapoptosis
spellingShingle Yulong Liu
Yulong Liu
Xinyue Wang
Xinyue Wang
Maomao Zhang
Maomao Zhang
Dan Cheng
Dan Cheng
Zhenpeng Zhu
Lan Ge
Xiaoyu Cheng
Mechanism of Ershen Zhenwu Decoction in ameliorating chronic heart failure via JNK/MAPK-regulated apoptosis: insights from network pharmacology and experimental validation
Frontiers in Cardiovascular Medicine
Ershen Zhenwu decoction
network pharmacology
molecular docking
JNK/MAPK
apoptosis
title Mechanism of Ershen Zhenwu Decoction in ameliorating chronic heart failure via JNK/MAPK-regulated apoptosis: insights from network pharmacology and experimental validation
title_full Mechanism of Ershen Zhenwu Decoction in ameliorating chronic heart failure via JNK/MAPK-regulated apoptosis: insights from network pharmacology and experimental validation
title_fullStr Mechanism of Ershen Zhenwu Decoction in ameliorating chronic heart failure via JNK/MAPK-regulated apoptosis: insights from network pharmacology and experimental validation
title_full_unstemmed Mechanism of Ershen Zhenwu Decoction in ameliorating chronic heart failure via JNK/MAPK-regulated apoptosis: insights from network pharmacology and experimental validation
title_short Mechanism of Ershen Zhenwu Decoction in ameliorating chronic heart failure via JNK/MAPK-regulated apoptosis: insights from network pharmacology and experimental validation
title_sort mechanism of ershen zhenwu decoction in ameliorating chronic heart failure via jnk mapk regulated apoptosis insights from network pharmacology and experimental validation
topic Ershen Zhenwu decoction
network pharmacology
molecular docking
JNK/MAPK
apoptosis
url https://www.frontiersin.org/articles/10.3389/fcvm.2025.1561963/full
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