Study On The Mechanism Of Qianyang Fengsui Pill In Treating Insomnia Based On Network Pharmacology

Objective: In this study,a network pharmacological approach was used to investigate the effective components and mechanism of action of Qianyang Fengsui Pill in the treatment of insomnia.Methods: In this study,we applied the Traditional Chinese Medicine Systematic Pharmacology Analysis Platform(TCMSP),BATMAN-TCM,specialized chemical databases and literature review,and searched and collected the five Chinese herbs in Qianyang Fengsui Pill by oral bioavailability(OB)≥ 30% and drug-like properties(DL)≥ 0.18.The active chemical components and the targets corresponding to the active chemical components were collected.The results were checked in the Uniprot database to obtain the effective target information of Qianyang Fengsui Pill;the target information of insomnia-related diseases was screened by the human genome annotation database(Gene Cards)and Drug Bank;the two groups of genes were mapped using the Venn diagram to obtain the intersecting target genes as the potential targets of this formula for the treatment of insomnia.The topological characteristics of the nodes in the network were evaluated by using Cytoscape’s cytohubba plug-in to calculate the parameters of degree centrality(DC),medial centrality(between),and protein interaction network(PPI).The target genes ranked in the top 10 of DC,BC and CC algorithms were intersected to obtain the core targets of Qianyang Fengsui Pill for insomnia.The top 15 protein level bar graphs according to Degree ranking were drawn using the Microbiology website;the intersecting genes were entered into the Metascape database for gene ontology(GO)function and Kyoto Encyclopedia of Genomes(KEGG)pathway enrichment analysis to obtain relevant biological processes and signaling pathways.We also used the Microbiology website to create bubble diagrams and triple histograms of the top 20 pathways according to the number of Gene in descending order,and the network visualization software Cytoscape 3.8.2 to map the biological network of the action of Qianyang Fengsui Pill in insomnia,i.e.,to build a "disease-drug-component-target-pathway" network.Molecular docking of core target proteins with key active ingredients was verified using Autodock and Pymol software.Results:1.The active ingredients and targets of Qianyang Fengsui Pill: Through various databases and literature search,we obtained 6 active ingredients of Aconiti Lateralis Radix Praeparata,10 of Amomum villosum,13 of phellodendri chinensis cortex,3 of Tortoise shell were extracted and 92 of Licorice,among which the important ingredients are quercetin,beta-sitosterol,Isocorypalmine,Kaempferol and(S)-Canadine.A total of 303 targets were obtained after de-weighting,including 18 active targets of Aconiti Lateralis Radix Praeparata,171 active targets of Amomum villosum,188 active targets of phellodendri chinensis cortex,87 active targets of Tortoise shell and 226 active targets of Licorice.2.Insomnia-related disease target genes: A total of 693 targets were screened through various databases.3.Intersection target protein interaction network: A total of 110 intersecting targets were obtained by taking the intersection.There are 109 nodes and 1057 edges in the protein interactions network,and the top 15 significant targets include IL-6,AKT1,TNF,IL1 B,CASP3,PTGS2,PPARG,VEGFA,APOE,CAT,CXCL8,ESR1,MMP9,NOS3,FOS,according to the topological characteristics.There are three core targets,namely APOE,IL-6,and AKT1.4.Enrichment analysis results: GO analysis yielded 1632 biological processes,100 cellular compositions,and 173 molecular functions.263 signaling pathways were derived from KEGG pathway enrichment library analysis.In BP,the main pathways are chemical synaptic transmission,cellular response to organic cyclic compounds,and cellular response to nitrogenous compounds.In CC,they are mainly synapses,dendrites,neuronal cell bodies,etc.In MF,mainly neurotransmitter receptor activity,G protein-coupled amine receptor activity,ion channel activity,etc.In KEGG,it is mainly neuroactive ligand-receptor interaction,calcium signaling pathway,serotonergic synapse,IL-17 signaling pathway,etc.5.Construction of "disease-drug-component-target-pathway" network: In the "disease-drug-component-target-pathway" network,the top 5 active components in terms of degree-value were quercetin,beta-sitosterol,tetrahydroafrican hexosine,kaempferol,and tetrahydroberberine,the top 5key targets were PTGS2,CALM1,ESR1,PPARG,and GSK3 B,and the top 5KEGG pathways were neuroactive ligand-receptor interaction,calcium signaling pathway,pathway in cancer,and AGE-RAGE signaling in diabetic complications.PPARG,GSK3 B,and the top 5 KEGG pathways were neuroactive ligand-receptor interaction,calcium signaling pathway,pathway in cancer,AGE-RAGE signaling pathway in diabetic complications,and fluid shear stress and atherosclerosis,respectively.It shows that the treatment of insomnia with Qianyang Fengsui Pill is a multi-component,multi-target and multi-pathway therapy.6.Molecular docking results: The molecular docking results showed that the five active ingredients,beta-sitosterol,quercetin,kaempferol,tetrahydroberberberine,and tetrahydroafrican hexylamine,were negative with the three target proteins,AKT1,APOE,and IL-6,and the binding force of AKT1 to all five compounds was less than-7.0 kcal-mol-1.The docking results of AKT1 and the five compounds were visualized,and the visualization revealed that AKT1 and the five compounds docking results were visualized,and through visualization,it can be found that the binding was mainly through hydrogen bonding,hydrophobic interaction and salt bridging interaction.It can be seen that the key chemical components of Qianyang Fengsui Pill for insomnia have strong binding to the core targets.Conclusion:1.Qianyang Fengsui pill plays a role in the treatment of insomnia through multi-component,multi-target and multi-channel.2.The mechanism of Qianyang Fengsui pill in the treatment of insomnia is closely related to quercetin,kaempferol,beta sitosterol,Isocorypalmine tetrandrine,(S)-Canadine and other active components.3.Qianyang Fengsui pill is mainly used to treat insomnia by affecting APOE,IL-6,AKT1,PTGS2,CALM1,ESR1,PPARG,GSK3 B,TNF,IL1 B,CASP3,VEGFA and other targets.It is speculated that Qianyang Fengsui pill is mainly used to treat insomnia by affecting neurotransmitters in the central nervous system.4.Qianyang Fengsui pill mainly regulates the interaction between neuroactive ligands and receptors,calcium signal transduction pathway,serotonin synapse,c AMP signal pathway,PI3 K Akt signal pathway,TNF signal transduction pathway,IL-17 signal transduction pathway and dopaminergic synapse pathway,so as to treat insomnia.5.Through molecular docking technology,it is preliminarily verified that the active components of Qianyang Fengsui pill are closely combined with the relevant targets of insomnia.