Objective Traditional Chinese medicine(TCM)constitutes a valuable cultural heritage and an important source of antitumor compounds.Poria(Poria cocos(Schw.)Wolf),the dried sclerotium of a polyporaceae fungus,was first ...Objective Traditional Chinese medicine(TCM)constitutes a valuable cultural heritage and an important source of antitumor compounds.Poria(Poria cocos(Schw.)Wolf),the dried sclerotium of a polyporaceae fungus,was first documented in Shennong’s Classic of Materia Medica and has been used therapeutically and dietarily in China for millennia.Traditionally recognized for its diuretic,spleen-tonifying,and sedative properties,modern pharmacological studies confirm that Poria exhibits antioxidant,anti-inflammatory,antibacterial,and antitumor activities.Pachymic acid(PA;a triterpenoid with the chemical structure 3β-acetyloxy-16α-hydroxy-lanosta-8,24(31)-dien-21-oic acid),isolated from Poria,is a principal bioactive constituent.Emerging evidence indicates PA exerts antitumor effects through multiple mechanisms,though these remain incompletely characterized.Neuroblastoma(NB),a highly malignant pediatric extracranial solid tumor accounting for 15%of childhood cancer deaths,urgently requires safer therapeutics due to the limitations of current treatments.Although PA shows multi-mechanistic antitumor potential,its efficacy against NB remains uncharacterized.This study systematically investigated the potential molecular targets and mechanisms underlying the anti-NB effects of PA by integrating network pharmacology-based target prediction with experimental validation of multi-target interactions through molecular docking,dynamic simulations,and in vitro assays,aimed to establish a novel perspective on PA’s antitumor activity and explore its potential clinical implications for NB treatment by integrating computational predictions with biological assays.Methods This study employed network pharmacology to identify potential targets of PA in NB,followed by validation using molecular docking,molecular dynamics(MD)simulations,MM/PBSA free energy analysis,RT-qPCR and Western blot experiments.Network pharmacology analysis included target screening via TCMSP,GeneCards,DisGeNET,SwissTargetPrediction,SuperPred,and PharmMapper.Subsequently,potential targets were predicted by intersecting the results from these databases via Venn analysis.Following target prediction,topological analysis was performed to identify key targets using Cytoscape software.Molecular docking was conducted using AutoDock Vina,with the binding pocket defined based on crystal structures.MD simulations were performed for 100 ns using GROMACS,and RMSD,RMSF,SASA,and hydrogen bonding dynamics were analyzed.MM/PBSA calculations were carried out to estimate the binding free energy of each protein-ligand complex.In vitro validation included RT-qPCR and Western blot,with GAPDH used as an internal control.Results The CCK-8 assay demonstrated a concentration-dependent inhibitory effect of PA on NB cell viability.GO analysis suggested that the anti-NB activity of PA might involve cellular response to chemical stress,vesicle lumen,and protein tyrosine kinase activity.KEGG pathway enrichment analysis suggested that the anti-NB activity of PA might involve the PI3K/AKT,MAPK,and Ras signaling pathways.Molecular docking and MD simulations revealed stable binding interactions between PA and the core target proteins AKT1,EGFR,SRC,and HSP90AA1.RT-qPCR and Western blot analyses further confirmed that PA treatment significantly decreased the mRNA and protein expression of AKT1,EGFR,and SRC while increasing the HSP90AA1 mRNA and protein levels.Conclusion It was suggested that PA may exert its anti-NB effects by inhibiting AKT1,EGFR,and SRC expression,potentially modulating the PI3K/AKT signaling pathway.These findings provide crucial evidence supporting PA’s development as a therapeutic candidate for NB.展开更多
BACKGROUND Pachymic acid(PA)is derived from Poria cocos.PA has a variety of pharmacological and inhibitory effects on various tumors.However,the mechanism of action of PA in gastric cancer(GC)remains unclear.AIM To in...BACKGROUND Pachymic acid(PA)is derived from Poria cocos.PA has a variety of pharmacological and inhibitory effects on various tumors.However,the mechanism of action of PA in gastric cancer(GC)remains unclear.AIM To investigate the mechanism of PA in treating GC via the combination of network pharmacology and experimental verification.METHODS The GeneCards and OMIM databases were used to derive the GC targets,while the Pharm Mapper database provided the PA targets.Utilizing the STRING database,a protein-protein interaction network was constructed and core targets were screened.The analyses of Gene Ontology,Kyoto Encyclopedia of Genes and Genomes(KEGG),and gene set enrichment analysis were conducted,and molecular docking and clinical correlation analyses were performed on the core targets.Ultimately,the network pharmacology findings were validated through in vitro cell assays,encompassing assessments of cell viability,apoptosis,cell cycle,cloning,and western blot analysis.RESULTS According to network pharmacology analysis,the core targets were screened,and the PI3K/AKT signaling pathway is likely to be the mechanism by which PA effectively treats GC,according to KEGG enrichment analysis.The experimental findings showed that PA could control PI3K/AKT signaling to prevent GC cell proliferation,induce apoptosis,and pause the cell cycle.CONCLUSION Network pharmacology demonstrated that PA could treat GC by controlling a variety of signaling pathways and acting on a variety of targets.This has also been supported by in vitro cell studies,which serve as benchmarks for further research.展开更多
Objective:To determine the inhibitory effects of pachymic acid on lung adenocarcinoma(LUAD)cells and elucidate its underlying mechanism.Methods:CCK-8,wound healing,Transwell,Western blot,tube formation,and immunofluor...Objective:To determine the inhibitory effects of pachymic acid on lung adenocarcinoma(LUAD)cells and elucidate its underlying mechanism.Methods:CCK-8,wound healing,Transwell,Western blot,tube formation,and immunofluorescence assays were carried out to measure the effects of various concentrations of pachymic acid on LUAD cell proliferation,metastasis,angiogenesis as well as autophagy.Subsequently,molecular docking technology was used to detect the potential targeted binding association between pachymic acid and protein tyrosine phosphatase 1B(PTP1B).Moreover,PTP1B was overexpressed in A549 cells to detect the specific mechanisms of pachymic acid.Results:Pachymic acid suppressed LUAD cell viability,metastasis as well as angiogenesis while inducing cell autophagy.It also targeted PTP1B and lowered PTP1B expression.However,PTP1B overexpression reversed the effects of pachymic acid on metastasis,angiogenesis,and autophagy as well as the expression of Wnt3a andβ-catenin in LUAD cells.Conclusions:Pachymic acid inhibits metastasis and angiogenesis,and promotes autophagy in LUAD cells by modulating the Wnt/β-catenin signaling pathway via targeting PTP1B.展开更多
Objective:Pulmonary hypertension(PH)is a severe pulmonary vascular disease that eventually leads to right ventricular failure and death.The purpose of this study was to investigate the mechanism by which pachymic acid...Objective:Pulmonary hypertension(PH)is a severe pulmonary vascular disease that eventually leads to right ventricular failure and death.The purpose of this study was to investigate the mechanism by which pachymic acid(PA)pretreatment affects PH and pulmonary vascular remodeling in rats.Methods:PH was induced via hypoxia exposure and administration of PA(5 mg/kg per day)in male Sprague-Dawley rats.Hemodynamic parameters were measured using a right ventricular floating catheter and pulmonary vascular morphometry was measured by hematoxylin-eosin(HE),a-SMA and Masson staining.MTT assays and EdU staining were used to detect cell proliferation,and apoptosis was analyzed by TUNEL staining.Western blotting and immunohistochemistry were used to detect the expression of proteins related to the Nrf2-Keapl-ARE pathway.展开更多
Background Pachymic acid （PA）, a natural triterpenoid, is known to significantly reduce cell proliferation and induce apoptosis in vitro through initiation of mitochondria dysfunction. However, its effect on immune ...Background Pachymic acid （PA）, a natural triterpenoid, is known to significantly reduce cell proliferation and induce apoptosis in vitro through initiation of mitochondria dysfunction. However, its effect on immune cells and anti-rejection following organ transplantation remains unknown. Methods In this study, we investigated PA as a treatment to control acute rejection occurred in rats which had accepted cardiac transplantation. We measured apoptosis of peripheral blood lymphocyte （PBLs）, and CD4^＋ lymphocyte, as well as the number of CD4^＋ and CD8^＋ lymphocytes and the effect of PA on acute rejection in rats 7 days after cardiac transplantation. Results PA treatment might decrease allograft rejection, protect PBLs from apoptosis, and reduce the percentage of CD8^＋ lymphocyte. PA neither regulated the number nor the apoptosis rate of CD4^＋ lymphocyte. Conclusions Our findings indicated that PA has an anti-apoptotic effect acting on PBLs through a novel mechanism involving stabilization of the PBLs mitochondrial transmembrane potential, an anti-rejection effect in rats after cardiac transplantation and an inhibiting effect to CD8^＋ lymphocyte.展开更多
文摘Objective Traditional Chinese medicine(TCM)constitutes a valuable cultural heritage and an important source of antitumor compounds.Poria(Poria cocos(Schw.)Wolf),the dried sclerotium of a polyporaceae fungus,was first documented in Shennong’s Classic of Materia Medica and has been used therapeutically and dietarily in China for millennia.Traditionally recognized for its diuretic,spleen-tonifying,and sedative properties,modern pharmacological studies confirm that Poria exhibits antioxidant,anti-inflammatory,antibacterial,and antitumor activities.Pachymic acid(PA;a triterpenoid with the chemical structure 3β-acetyloxy-16α-hydroxy-lanosta-8,24(31)-dien-21-oic acid),isolated from Poria,is a principal bioactive constituent.Emerging evidence indicates PA exerts antitumor effects through multiple mechanisms,though these remain incompletely characterized.Neuroblastoma(NB),a highly malignant pediatric extracranial solid tumor accounting for 15%of childhood cancer deaths,urgently requires safer therapeutics due to the limitations of current treatments.Although PA shows multi-mechanistic antitumor potential,its efficacy against NB remains uncharacterized.This study systematically investigated the potential molecular targets and mechanisms underlying the anti-NB effects of PA by integrating network pharmacology-based target prediction with experimental validation of multi-target interactions through molecular docking,dynamic simulations,and in vitro assays,aimed to establish a novel perspective on PA’s antitumor activity and explore its potential clinical implications for NB treatment by integrating computational predictions with biological assays.Methods This study employed network pharmacology to identify potential targets of PA in NB,followed by validation using molecular docking,molecular dynamics(MD)simulations,MM/PBSA free energy analysis,RT-qPCR and Western blot experiments.Network pharmacology analysis included target screening via TCMSP,GeneCards,DisGeNET,SwissTargetPrediction,SuperPred,and PharmMapper.Subsequently,potential targets were predicted by intersecting the results from these databases via Venn analysis.Following target prediction,topological analysis was performed to identify key targets using Cytoscape software.Molecular docking was conducted using AutoDock Vina,with the binding pocket defined based on crystal structures.MD simulations were performed for 100 ns using GROMACS,and RMSD,RMSF,SASA,and hydrogen bonding dynamics were analyzed.MM/PBSA calculations were carried out to estimate the binding free energy of each protein-ligand complex.In vitro validation included RT-qPCR and Western blot,with GAPDH used as an internal control.Results The CCK-8 assay demonstrated a concentration-dependent inhibitory effect of PA on NB cell viability.GO analysis suggested that the anti-NB activity of PA might involve cellular response to chemical stress,vesicle lumen,and protein tyrosine kinase activity.KEGG pathway enrichment analysis suggested that the anti-NB activity of PA might involve the PI3K/AKT,MAPK,and Ras signaling pathways.Molecular docking and MD simulations revealed stable binding interactions between PA and the core target proteins AKT1,EGFR,SRC,and HSP90AA1.RT-qPCR and Western blot analyses further confirmed that PA treatment significantly decreased the mRNA and protein expression of AKT1,EGFR,and SRC while increasing the HSP90AA1 mRNA and protein levels.Conclusion It was suggested that PA may exert its anti-NB effects by inhibiting AKT1,EGFR,and SRC expression,potentially modulating the PI3K/AKT signaling pathway.These findings provide crucial evidence supporting PA’s development as a therapeutic candidate for NB.
基金Supported by Ningxia Science and Technology Benefiting People Program,No.2022CMG03064National Natural Science Foundation of China,No.82260879Ningxia Natural Science Foundation,No.2022AAC03144 and 2022AAC02039.
文摘BACKGROUND Pachymic acid(PA)is derived from Poria cocos.PA has a variety of pharmacological and inhibitory effects on various tumors.However,the mechanism of action of PA in gastric cancer(GC)remains unclear.AIM To investigate the mechanism of PA in treating GC via the combination of network pharmacology and experimental verification.METHODS The GeneCards and OMIM databases were used to derive the GC targets,while the Pharm Mapper database provided the PA targets.Utilizing the STRING database,a protein-protein interaction network was constructed and core targets were screened.The analyses of Gene Ontology,Kyoto Encyclopedia of Genes and Genomes(KEGG),and gene set enrichment analysis were conducted,and molecular docking and clinical correlation analyses were performed on the core targets.Ultimately,the network pharmacology findings were validated through in vitro cell assays,encompassing assessments of cell viability,apoptosis,cell cycle,cloning,and western blot analysis.RESULTS According to network pharmacology analysis,the core targets were screened,and the PI3K/AKT signaling pathway is likely to be the mechanism by which PA effectively treats GC,according to KEGG enrichment analysis.The experimental findings showed that PA could control PI3K/AKT signaling to prevent GC cell proliferation,induce apoptosis,and pause the cell cycle.CONCLUSION Network pharmacology demonstrated that PA could treat GC by controlling a variety of signaling pathways and acting on a variety of targets.This has also been supported by in vitro cell studies,which serve as benchmarks for further research.
基金supported by the Zhejiang Province Traditional Chinese Medicine Health Science and Technology Program(2023ZL570).
文摘Objective:To determine the inhibitory effects of pachymic acid on lung adenocarcinoma(LUAD)cells and elucidate its underlying mechanism.Methods:CCK-8,wound healing,Transwell,Western blot,tube formation,and immunofluorescence assays were carried out to measure the effects of various concentrations of pachymic acid on LUAD cell proliferation,metastasis,angiogenesis as well as autophagy.Subsequently,molecular docking technology was used to detect the potential targeted binding association between pachymic acid and protein tyrosine phosphatase 1B(PTP1B).Moreover,PTP1B was overexpressed in A549 cells to detect the specific mechanisms of pachymic acid.Results:Pachymic acid suppressed LUAD cell viability,metastasis as well as angiogenesis while inducing cell autophagy.It also targeted PTP1B and lowered PTP1B expression.However,PTP1B overexpression reversed the effects of pachymic acid on metastasis,angiogenesis,and autophagy as well as the expression of Wnt3a andβ-catenin in LUAD cells.Conclusions:Pachymic acid inhibits metastasis and angiogenesis,and promotes autophagy in LUAD cells by modulating the Wnt/β-catenin signaling pathway via targeting PTP1B.
基金This project was supported by the Natural Science Foundation of Hubei Province(No.2017CFB769).
文摘Objective:Pulmonary hypertension(PH)is a severe pulmonary vascular disease that eventually leads to right ventricular failure and death.The purpose of this study was to investigate the mechanism by which pachymic acid(PA)pretreatment affects PH and pulmonary vascular remodeling in rats.Methods:PH was induced via hypoxia exposure and administration of PA(5 mg/kg per day)in male Sprague-Dawley rats.Hemodynamic parameters were measured using a right ventricular floating catheter and pulmonary vascular morphometry was measured by hematoxylin-eosin(HE),a-SMA and Masson staining.MTT assays and EdU staining were used to detect cell proliferation,and apoptosis was analyzed by TUNEL staining.Western blotting and immunohistochemistry were used to detect the expression of proteins related to the Nrf2-Keapl-ARE pathway.
基金The work was supported by a grant from the Molecular Biology Laboratory of the Second Hospital Affiliated to Harbin Medical University （Harbin, China） for building animal model, cellular extraction and staining, the Pathological Research Center of the Harbin Medical University （Harbin, China） for producing and assessment tissular section and the Dairy Science Center of Northeast Agricultural University （Harbin, China） for flow cytometry analysis. This studying was supported by a grant from National Nature Science Foundation of China （No. 30671975）.
文摘Background Pachymic acid （PA）, a natural triterpenoid, is known to significantly reduce cell proliferation and induce apoptosis in vitro through initiation of mitochondria dysfunction. However, its effect on immune cells and anti-rejection following organ transplantation remains unknown. Methods In this study, we investigated PA as a treatment to control acute rejection occurred in rats which had accepted cardiac transplantation. We measured apoptosis of peripheral blood lymphocyte （PBLs）, and CD4^＋ lymphocyte, as well as the number of CD4^＋ and CD8^＋ lymphocytes and the effect of PA on acute rejection in rats 7 days after cardiac transplantation. Results PA treatment might decrease allograft rejection, protect PBLs from apoptosis, and reduce the percentage of CD8^＋ lymphocyte. PA neither regulated the number nor the apoptosis rate of CD4^＋ lymphocyte. Conclusions Our findings indicated that PA has an anti-apoptotic effect acting on PBLs through a novel mechanism involving stabilization of the PBLs mitochondrial transmembrane potential, an anti-rejection effect in rats after cardiac transplantation and an inhibiting effect to CD8^＋ lymphocyte.
