Exploring the Mechanism of Action of Glyasperin A in Intervening Menopause Based on Network Pharmacology and Molecular Docking Technology

[Objectives]To investigate the mechanism of action of glyasperin A(GAA)in intervening menopause using network pharmacology and molecular docking technology.[Methods]All target names of the active ingredients were screened using TCMSP,3D model molecules converted into SMILES online tool,Swiss target prediction and literature search.The relevant target genes corresponding to menopause were identified using the Genecards database.Venn 2.1.0 was then used to generate the corresponding Venn diagram.Finally,the protein-protein interaction(PPI)network was constructed using Cytoscape 3.9.1 software.The core targets that were screened underwent enrichment and analysis using the Gene Ontology(GO)biological process and KEGG pathways with the assistance of the DAVID database and bioinformatics.The molecular docking was then verified using AutoDock and Pymol software on the core targets.[Results]This study screened 100 target genes of active ingredients.In the PPI network,ESR1 and AKT1 were found to have a higher degree.The GO and KEGG enrichment analyses revealed that the biological processes primarily involved platelet activation,regulation of circadian rhythms,and regulation of mRNA stability.The signalling pathways included hepatitis B,cytotoxicity,and gastric cancer.The molecular docking results indicated that the key active ingredients and proteins bound well,as evidenced by their small binding energies.[Conclusions]Using a systematic network pharmacology approach,this study predicts the basic pharmacological effects and potential mechanisms of GAA in intervening menopause,which provides a foundation for further research on its pharmacological mechanisms.

Study on the Mechanism of Action of Glyasperin A in the Treatment of Atherosclerosis Based on Network Pharmacology and Molecular Docking

[Objectives] This study was conducted to investigate the mechanism of action of glyasperin A in the treatment of atherosclerosis using a network pharmacology approach. [Methods] Targets related to atherosclerosis were searched in GeneCards database. An active ingredient-disease-target network was constructed by Cytoscape 3.7.1. A target protein interaction network was constructed by String database. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed on the DAVID database. [Results] Glyasperin A acted on 36 atherosclerosis-related targets, and the biofunctional and pathway enrichment analyses showed that it was mainly involved in response to xenobiotic stimulus, drug transport across blood-brain barrier, lipid oxidation, barrier, and lipid oxidation, etc. The results showed that glyasperin A acted on 36 atherosclerosis-related targets. The biofunctional and pathway enrichment analyses showed that it was mainly involved in response to xenobiotic stimulus, drug transport across blood-brain barrier, lipid oxidation, positive regulation of protein localization to nucleus, and hepoxilin biosynthetic process, and it played an anti-fatigue role through signal pathways such as serotonergic synapse, efferocytosis, arachidonic acid metabolism, chemical carcinogenesis-receptor activation and platelet activation. [Conclusions] Glyasperin A has multi-target and multi-pathway effects in the treatment of atherosclerosis. This study provides reference for further research on glyasperin A in the treatment of atherosclerosis.

Therapeutic Effects and Potential Mechanisms of Glyasperin A against Myocardial Ischemia Based on Network Pharmacology and Molecular Docking

[Objectives]To explore the therapeutic effects and potential mechanisms of Glyasperin A(GAA)on myocardial ischemia(MI)based on network pharmacology and molecular docking.[Methods]The molecular structure of GAA was downloaded from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),and all targets of GAA were predicted by converting 3D model molecules into SMILES online tool and Swiss target prediction.Genecards database and DisGeNET database were used to find the targets related to MI,and then Venny 2.1.0 was used to generate the corresponding Wayne diagram,and then Cytoscape 3.9.1 software was used to construct the protein-protein interaction(PPI)network.With the help of DAVID database and Microbiology,the selected core targets were enriched and analyzed by gene ontology(GO),biological process(BP),and Kyoto Encyclopedia of Genes and Genomes(KEGG),and then the molecular docking between GAA and core targets was verified by AutoDock and Pymol software.[Results]A total of 1883 MI targets were screened,and in the protein-protein interaction network,AKT1,PTGS2,PPARG,ESR1,GSK3B were the proteins with higher values.Gene ontology and KEEG enrichment analysis showed that the biological processes involved mainly included inflammatory response,negative regulation of gene expression,and response to exogenous stimuli.Signaling pathways mainly include IL-17 signaling pathway,HIF-1 signaling pathway,and so on.The results of molecular docking showed that the binding energy of GAA and core protein was less than-5 Kcal/mol in four groups.These indicated that GAA with good binding had a certain therapeutic effect on myocardial ischemia.[Conclusions]Based on the systematic network pharmacology method,this study predicts the basic pharmacological effects and potential mechanisms of GAA in the treatment of MI,and reveals that GAA may treat MI through multiple targets and signaling pathways.It is expected to provide a basis for further study of its pharmacological mechanisms.

Therapeutic Effect and Mechanism of Jipei Dilong Ointment on Acute Soft Tissue Injury in Rats

[Objectives]This study was conducted to observe the therapeutic effect of Jipei Dilong Ointment on rats with acute soft tissue injury caused by heavy objects and to explore its action mechanism.[Methods]Thirty six rats were randomly divided into six groups(control group,model group,high-dose Jipei Dilong Ointment group(JP-H),medium-dose Jipei Dilong Ointment group(JP-M),low-dose Jipei Dilong Ointment group(JP-L)and diclofenac group).Except for the Control group,other groups were subjected to modeling of acute soft tissue injury by the weight impact method.All administration was performed once a day for nine consecutive days.The local appearance score and activity disorder score were determined after soft tissue injury in rats.HE staining was used to detect the pathological changes of injured soft tissues in rats.RT-PCR was used to detect the relative mRNA expressions of Bax,Bcl-2,MMP-9 and TIMP-1 in injured soft tissues of rats.Western Blot was used to detect the protein expressions of MMP-9,TIMP-1,TLR4,MyD88 and NF-κB p65 in injured soft tissues of rats.Results were statistically analyzed.[Results]Compared with the model group,Jipei Dilong Ointment could significantly improve the appearance symptoms such as swelling and ecchymosis in the injured area and the movement function of the affected limb(P<0.05).It could also improve the infiltration of inflammatory cells and widening of the intermuscular space caused by injury.Among them,the JP-H group and the diclofenac group had more significant curative effects.After 9 d of administration,each administration group could significantly up-regulate the ratio of Bcl-2/Bax mRNA expression level(P<0.05 or P<0.01),and the ratio of MMP-9/TIMP-1 mRNA expression level showed a downward trend(P>0.05).The expression level of NF-κB p65 protein in each administration group was significantly decreased(P<0.01).The protein expression levels of TLR4 and MyD88 and the ratio of MMP-9/TIMP-1 protein expression level in each administration group decreased to varying degrees.Among them,the JP-H group and diclofenac group significantly decreased(P<0.05).[Conclusions]Jipei Dilong Ointment has the functions of relieving pain,swelling and inflammation.It could improve the local appearance,functional activity and tissue morphology of affected limbs in rats,and has a therapeutic effect on acute soft tissue injury in rats.Its mechanism of action might be related to the inhibition of TLR4/MyD88/NF-κB p65 signaling pathway and the regulation of Bcl-2/Bax and MMP-9/TIMP-1 balance.

Study on the Mechanism of Pseudostellariae Radix in Regulating Angiogenesis Based on Network Pharmacology and a Dual-screening System

[Objectives]This study was conducted to clarify the action mechanism of Pseudostellariae Radix in regulating angiogenesis by using network pharmacology and a dual-screening system,and to provide a basis for its clinical treatment of cardiovascular diseases.[Methods]The TCMSP database was used for preliminary screening to obtain the active compounds of Pseudostellariae Radix and the protein targets of its action.GeneCards and OMIM databases were used to search for targets related to angiogenesis.Cytoscape 3.9.1 was used to construct a drug-target network and protein interaction network of Pseudostellariae Radix in angiogenesis.The GO enrichment analysis and KEGG pathway analysis of the targets of Pseudostellariae Radix in angiogenesis were carried out on Metascape platform.The effects of the screened active compounds were verified using a dual-screening system.[Results]Six active components of Pseudostellariae Radix,luteolin,acetin,beta-sitosterol,linarin,schottenol and 1-monolinolein,were screened by TCMSP database;and the six active components were predicted with 78 common target proteins related to angiogenesis,of which 19 were core targets.Pseudostellariae Radix mainly intervened in angiogenesis through domain specific binding,ubiquitin-like protein ligase binding,kinase binding and other molecular functions to regulate biological processes such as membrane microdomain,plasma membrane raft and caveola.The results of KEGG enrichment indicated that pathways in cancer,lipid and atherosclerosis,hepatitis B,apoptosis,toxoplasmosis and other key pathways might be the mechanism for the intervention of angiogenesis.The results of the dual-screening system showed that luteolin,acacetin,beta-sitosterol and linarin protected HUVECs and promoted zebrafish angiogenesis.[Conclusions]This study preliminarily demonstrated that luteolin,acacetin,beta-sitosterol and linarin could intervene in angiogenesis through multiple targets and multiple pathways,providing ideas and a scientific basis for the treatment of cardiovascular diseases.

Protective Effect and Molecular Mechanism of Shui People's Classic Prescription Jipei Dilong Ointment on Osteoarthritis in Rats

[Objectives]To explore the protective effect and molecular mechanism of Shui People s Classic Prescription Jipei Dilong Ointment on knee osteoarthritis in rats.[Methods]72 SPF male SD rats were divided into control group,model group,Jipei Dilong Ointment high,medium and low dose groups,and positive drug Diclofenac group,with 12 rats in each group.Except the control group,all other groups were injected with 0.2 mL of 2%papain and 0.3%L-cysteine mixture into knee joint cavity to establish osteoarthritis model,while the control group was injected with the same amount of normal saline.The Lequesne MG score was used to determine the success of the model.After successful modeling,external administration was given for 4 weeks.The histopathological changes in articular cartilage and synovium were observed by HE staining;the levels of TNF-α,IL-1βand COX-2 in serum were detected by ELISA,and the relative expression of MMP-9 and TIMP-1 mRNA in cartilage was detected by qRT-PCR;the relative expression of MMP-9,TIMP-1,TLR4,MyD88 and NF-κB in rat cartilage was detected by Western-blot.[Results]Compared with the model group,Jipei Dilong Ointment could significantly reduce the Lequesne MG score and Mankin s score of arthritic rats(P<0.01);significantly improve the pathological changes in articular cartilage and synovium,reduce tissue edema,necrosis,inflammatory cell infiltration and fibrous tissue proliferation;reduce the expression of MMP-9 mRNA(P<0.01)in different degrees and increase the expression of TIMP-1 mRNA(P<0.01);reduce the relative expression of MMP-9,TLR4,MyD88 and NF-κB protein in different degrees(P<0.01),and significantly increase the relative expression of TIMP-1 protein in cartilage(P<0.01).[Conclusions]Jipei Dilong Ointment can improve the joint injury of osteoarthritis rats,and the mechanism may be related to the regulation of TLR4/MyD88/NF-κB signaling pathway and the ratio of MMP-9/TIMP-1.

Study on the Molecular Mechanism of Danggui Buxue Decoction in Intervention of Perimenopausal Syndrome Based on Network Pharmacology and Molecular Docking

[Objectives]This study was conducted to explore the intervention mechanism of Danggui Buxue Decoction in perimenopausal syndrome based on network pharmacology and molecular docking.[Methods]The chemical components and targets of Danggui Buxue Decoction were acquired through the TCMSP database,and the main targets of perimenopausal syndrome were obtained through the GeneCards database.The component targets and disease targets were intersected,and combining with active components and Chinese herbs in the decoction,a traditional Chinese medicine-component-target network was constructed using Cytoscape 3.7.1 software.The STRING platform was employed for protein-protein interaction analysis.The DAVID analysis platform was used to conduct target GO and KEGG enrichment analysis,so as to predict the action mechanism Danggui Buxue Decoction.Finally,an active component-disease target-signal pathway network diagram was constructed.[Results]Twenty two components in Danggui Buxue Decoction related to perimenopausal syndrome and 120 corresponding targets were obtained,including active components such as 1,7-dihydroxy-3,9-dimquercetin and kaempferol,and key targets such as TNF,ESR1 and PPARG.The results of GO analysis and KEEG analysis indicated that Danggui Buxue Decoction might regulate the transcription of RNA polymerase II promoter,DNA templating,gene expression,signal transduction,hypoxia response and other biological processes by regulating multiple signal pathways such as chemical carcinogenesis-receptor activation,cancer pathways,lipid and atherosclerosis,tryptophan metabolism,malaria,steroid hormone biosynthesis and chemical carcinogenesis-DNA adduct.[Conclusions]Danggui Buxue Decoction intervenes in perimenopausal syndrome through multiple components,targets and pathways,providing a basis for elucidating the intervention mechanism of Danggui Buxue Decoction and expanding its clinical application.