Material basis and pharmacodynamic mechanism of YangshenDingzhi granules in the intervention of viral pneumonia:Based on serum pharmacochemistry and network pharmacology

Background:YangshenDingzhi granules(YSDZ)are clinically effective in preventing and treating COVID-19.The present study elucidates the underlying mechanism of YSDZ intervention in viral pneumonia by employing serum pharmacochemistry and network pharmacology.Methods:The chemical constituents of YSDZ in the blood were examined using ultraperformance liquid chromatography-quadrupole/orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS).Potential protein targets were obtained from the SwissTargetPrediction database,and the target genes associated with viral pneumonia were identified using GeneCards,DisGeNET,and Online Mendelian Inheritance in Man(OMIM)databases.The intersection of blood component-related targets and disease-related targets was determined using Venny 2.1.Protein-protein interaction networks were constructed using the STRING database.The Metascape database was employed to perform enrichment analyses of Gene Ontology(GO)functions and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathways for the targets,while the Cytoscape 3.9.1 software was utilized to construct drug-component-disease-target-pathway networks.Further,in vitro and in vivo experiments were performed to establish the therapeutic effectiveness of YSDZ against viral pneumonia.Results:Fifteen compounds and 124 targets linked to viral pneumonia were detected in serum.Among these,MAPK1,MAPK3,AKT1,EGFR,and TNF play significant roles.In vitro tests revealed that the medicated serum suppressed the replication of H1N1,RSV,and SARS-CoV-2 replicon.Further,in vivo testing analysis shows that YSDZ decreases the viral load in the lungs of mice infected with RSV and H1N1.Conclusion:The chemical constituents of YSDZ in the blood may elicit therapeutic effects against viral pneumonia by targeting multiple proteins and pathways.

Preparation of Ephedra houttuynia Granule and Its Therapeutic Mechanism of Anti-Mycoplasma gallisepticum Infection Based on Network Pharmacology

The use of Chinese herbal medicines can replace antibiotics that cause drug-resistance problems,which are currently necessary for disease control.In this paper,a traditional Chinese medicine compound named Ephedra houttuynia granule for the treatment of Mycoplasma galliscepticum(MG)infection was prepared.Furthermore,its action mechanism was explored through network pharmacology.The optimal extraction and granulation processes of the compound were determined by high performance liquid chromatography(HPLC)method and L9 orthogonal test,and in the treatment experiment,Ephedra houttuynia granule showed a significant therapeutic effect on MG infection.In the study of network pharmacology,the results showed that the core targets of Ephedra houttuynia granule against MG infection were vascular endothelial growth factor(VEGFA),fos proto-oncogene(FOS),prepro-coagulation factor II(F2),etc.,the gene ontology/kyoto encyclopedia of genes and genomes(GO/KEGG)analysis results indicated that the signaling pathways of neuroactive ligand receptor interaction,cAMP,IL-17,T cell receptor,and tumor necrosis factor(TNF)might involve in anti-MG infection.In conclusion,this study would provide a new idea for elucidating the action mechanism of other diseases in veterinary clinic,which had a certain guiding significance.

To explore the mechanism of Fuyang Jiebiao granules against viral pneumonia based on network pharmacology and pharmacodynamics

Objective:To investigate the mechanism of Fuyang Jiebiao granule(FYJBKL)in the treatment of viral pneumonia.Methods:Firstly,a network model was constructed using network pharmacology to study the target expression sites of FYJBKL viral pneumonia,so as to determine the main targets and important signal transduction pathways for the treatment of viral pneumonia.Secondly,the main components of the drug and the main target are docked.Then,the fever,sweating and inflammation rat models were established to explore the antipyretic,sweating and anti-inflammatory mechanisms of FYJBKL.Finally,the contents of IL-17,IL-1β,TNF-αand IL-6 in blood samples of rats were analyzed by ELISA method,and the morphological changes of lung tissue were observed by HE staining.Results:Quercetin,luteolin,kaempferol,etc.,and the main mechanism targets are IL-17,IL-1β,TNF-α,IL-6 and so on.Thirty signal pathways were identified by KEGG enrichment analysis,including interleukin-17 signaling pathway(IL-17 signaling pathway),human cytomegalovirus infection pathway(human cytomegalovirus infection),Kaposi's sarcoma associated herpesvirus infection pathway(Kaposi's sarcoma-as-sociated herpesvirus infection)and so on.After the study of molecular docking,we found that the contact efficiency between active substances and possible key targets is good.The high and middle concentration groups of FYJBKL significantly decreased the expression of IL-17,IL-1β,TNF-αand IL-6 in the blood of rats with inflammation(P<0.05).FYJBKL significantly reduced the foot swelling induced by egg white and inhibited the increase of body temperature induced by yeast in rats(P<0.05).HE staining showed that FYJBKL improved pulmonary fibrosis and inflammatory exudation to varying degrees.Conclusion:The effects of FuyangJiebiao granules on the related signal pathways of anti-virus,anti-immune and anti-inflammation as well as biological and cellular processes may be caused by the binding of quercetin,luteolin,kaempferol and other active ingredients to their shared targets.Fuyang Jiebiao granules can improve the related symptoms caused by viral pneumonia,and its mechanism may be related to the activities of TNF,IL-17,IL-6 and other related channels,which are multiple targets of inflammation regulation.

Multicenter Clinical Randomized Controlled Trial and Network Pharmacology Analysis of Zhenzhu Qingyuan Granules for the Treatment of Gastroesophageal Reflux Disease

Objective To evaluate the clinical efficacy and safety of Zhenzhu Qingyuan Granules through a clinical randomized controlled trial and to analyze the potential action targets and pathways of this formula using network pharmacology.Methods Patients with gastroesophageal reflux disease(GERD)of liver–stomach stagnant heat pattern who met the inclusion and exclusion criteria were randomly divided into the control group and the observation group.The control group received oral rabeprazole,whereas the observation group were given Zhenzhu Qingyuan Granules in addition to the rabeprazole.The treatment duration was 8 weeks.Clinical efficacy was observed in both groups after 8 weeks.Network pharmacology was used to analyze the action targets of ZhenzhuQingyuanGranules and the genes related to GERD,and core targets were inferred.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted to explore the potential mechanisms of this formula.Results The clinical research results showed that the total effective rate in the treatment group was 92.68%,compared with 70.00%in the control group,with a statistically significant difference(p<0.05).After treatment,both Chinese medicine syndrome score and endoscopic score improved in both groups compared with before treatment(p<0.05),and the treatment group showed greater improvement than the control group(p<0.05).Network pharmacology identified effective components of Zhenzhu Qingyuan Granules for treating GERD,including quercetin,luteolin,andβ-sitosterol,with potential action targets such as tumor protein 53(TP53),protein kinase B(AKT1),and tumor necrosis factor.Conclusion Zhenzhu Qingyuan Granules can significantly improve clinical symptoms in patients with GERD of liver–stomach stagnated heat pattern,enhance clinical efficacy,and have high safety.This formula may exert therapeutic effects through multiple targets and pathways.

Mechanism of AiTongXiao granule in the treatment of hepatocellular carcinoma based on network pharmacology and rat transplanted liver cancer model

Objective:To investigate the mechanism of action and material basis of AiTongXiao granule in the treatment of hepatocellular carcinoma(HCC)based on network pharmacology and transplanted liver cancer rat model.Methods:TCMSP database was used to screen out effective components and its corresponding potential pharmaceutical targets,and databases including Gene Cards,OMIM,Drugbank and TTD were further used to collect HCC-related drug targets.The intersecting targets were obtained by mapping the drug and disease targets.The component-targets network was constructed and visualized by Cytoscape 3.8.2 software.Protein-protein interaction(PPI)network was built by STRING online platform,and the topological relationship and core targets was analyzed and screened by using CytoNCA software.In addition,Metascape database was used to perform gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis of the core targets.At last,rat liver transplanted liver cancer model was established by using Walker-256 cell line and treated by AiTongXiao granule for 15 days.Western blot was used to further compare the expression levels of AKT,pAKT,p53,p-p53,ERK1/2 and ERK1/2 in the tumor between treatment group and the control group.Results:257 active components were obtained from AiTongXiao granule,corresponding to 294 drug targets.Meanwhile,233 of the 7993 HCC disease targets were screened out between AiTongXiao granule drug and HCC disease targets.11 core targets including AKT1,IL6,TP53,MAPK3,TNF,JUN,CASP3,MAPK1,MYC,PTGS2,MMP9 were further obtained by median screening.GO and KEGG analysis results showed that these core targets enriched to HBV,TNF and cancer related pathways.The rat transplanted liver cancer model results indicated significant down regulation for AKT,p-AKT,pERK1/2,and significant up regulation of p-p53 after AiTongXiao granule treatment(P<0.05).Conclusion:AiTongXiao granule could act to multiple cancer related pathways,and AKT,p53 and ERK1/2 were validated to be regulated by ATXF in rat model.The mechanism may be through the regulation of the above signaling pathways to exert anti-liver cancer effect.

Investigation on the Material Basis of Sijicao Granules in Treating Eczema Based on Network Pharmacology

[Objectives]To explore the pharmacodynamic material basis of Sijicao granules for the treatment of eczema through chemical composition-network pharmacology.[Methods]First of all,the chemical constituents of Polygonum capitatum and Plantago asiatica from Sijicao granules were collected,and the relevant target information of the constituents was collected by TCMSP,PubChem,DisGeNET,GeneCards and STRING databases.Furthermore,Cytoscape 3.8.2 software was used to construct the chemical compounds-target network map of Sijicao granules.Finally,STRING database was used for PPI protein network analysis,GO functional enrichment analysis and KEGG pathway enrichment analysis of core targets,and molecular docking between core constituents and protein targets was also performed.[Results]30 constituents,including quercetin,kaempferol,luteolin,ellagic acid and gallic acid,were discovered to be the key effective compounds of Sijicao granules in the treatment of eczema.And its core action protein targets were PTGS2,NOS2,AKT1,TP53,IL6,HMOX1.What s more,through GO functional enrichment analysis of biological process(BP),cell component(CC),molecular function(MF)analysis and KEGG pathway enrichment analysis,the main pathways of action of Sijicao granules for the treatment of eczema including IL-17 signaling pathway,T cell receptor signaling pathway,cancer signaling pathway,TNF signaling pathway and Relaxin signaling pathway.In addition,molecular docking results displayed that the primary active constituents quercetin,kaempferol and luteolin were well combined with the core protein targets AKT1 and IL6.[Conclusions]Sijicao granules could play an important role for the treatment of eczema through multi-component,multi-target,multi-pathway and their interaction.

Acute and subchronic toxicity as well as evaluation of safety pharmacology of modified pulsatilla granules

The present study investigated acute and subchronic toxicity and safety pharmacology of modified pulsatilla granules（MPG）to provide a basis for a comprehensive understanding of MPG toxicity.The results of acute toxicity testing showed that the median lethal dose of MPG was more than 5 000 mg kg^-1,suggesting that MPG was considered as practically non-toxic.The subchronic toxicity study for 30 days was conducted by daily oral administration at doses of 375,750 and 1 500 mg kg^-1 in Sprague-Dawley rats.The results of subchronic toxicity study showed that the body weight and relative organ weight were not significantly changed by administration of MPG.The clinical chemistry study showed that MPG could induce kidney and liver damages.In histopathological,mild lesions in liver and kidney were also observed,suggesting that the liver and kidney might be potential target organs of MPG.In the safety pharmacology study,MPG did not exhibited any side effects to rats in cardiovascular system,respiratory system and central nervous system.These results suggested that MPG could be considered safe for veterinary use.

A Network Pharmacology Study to Uncover the Multiple Molecular Mechanism of the Chinese Patent Medicine Toujiequwen Granules in the Treatment of Corona Virus Disease 2019(COVID-19)

Since the outbreak of the novel corona virus disease 2019(COVID-19)at the end of 2019,specific antiviral drugs have been lacking.A Chinese patent medicine Toujiequwen granules has been promoted in the treatment of COVID-19.The present study was designed to reveal the molecular mechanism of Toujiequwen granules against COVID-19.A network pharmacological method was applied to screen the main active ingredients of Toujiequwen granules.Network analysis of 149 active ingredients and 330 drug targets showed the most active ingredient interacting with many drug targets is quercetin.Drug targets most ffected by the active ingredients were PTGS2,PTGS1,and DPP4.Drug target disease enrichment analysis showed drug targets were significantly enriched in cardiovascular diseases and digestive tract diseases.An"active ingredient-target-disease"network showed that 57 active ingredients from Toujiequwen granules interacted with 15 key targets of COVID-19.There were 53 ingredients that could act on DPP4,suggesting that DPP4 may become a potential new key target for the treatment of COVID-19.GO analysis results showed that key targets were mainly enriched in the cellular response to lipopolysaccharide,cytokine activity and other functions.KEGG analysis showed they were mainly concentrated in viral protein interaction with cytokine and cytokine receptors and endocrine resistance pathway.The evidence suggests that Toujiequwen granules might play an effective role by improving the symptoms of underlying diseases in patients with COVID-19 and multi-target interventions against mutiple signaling pathways related to the pathogenesis of COVID-19.

Network pharmacology deciphering multiple mechanisms of volatiles of Wendan granule for treatment of senile dementia

OBJECTIVE To explore the mechanisms of the volatiles of Wendan granule for the treatment of senile dementia,network pharmacology method integrating absorption,distribution,metab.olism,and excretion(ADME) screening,target fishing,network constructing,pathway analyzing,and correlated diseases prediction was applied.METHODS Twelve small molecular compounds of WDG were selected as the objects from 74 volatiles with the relative abundances above 2%,and their ADME parameters were collected from Traditional Chinese Medicine Systems Pharmacology platform(TCMSP),and then the corresponding targets,genes,pathways and diseases were predicted according to the data provided by TCMSP,DrugBank,Uniport and the Database for Annotation,Visualization and Integrated Discovery(DAVID).The related pathways and correlation analysis were explored by the Kyoto Encyclo.pedia and Genomes(KEGG) database.Finally,the networks of compound-target,target-pathway and pathway-disease of WDG were constructed by Cytoscape software.RESULTS Twelve compounds interacted with 49 targets,of which top three targets were Gamma-aminobutyric acid receptor subunit alpha-1(GABRA1),Prostaglandin G/H synthase 2(PGHS-2) and Sodium-dependent noradrenaline transporter.Interestingly,these targets were highly associated with depression,insomnia and Alzheimer′s disease that mainly corresponded to mental and emotional illnesses.CONCLUSION The integrated network pharmacology method provides precise probe to illuminate the molecular mechanisms of volatiles of WDG for relieving senile dementia related syndromes,which will also facilitate the application of traditional Chinese medicine in modern medicine,as well as follow-up studies such as upgrading the quality stan.dard of clinical medicine and novel drug development.

Network pharmacology-based prediction and verification of the mechanism for Bushen Chengyun granule on low endometrial receptivity

Objective:Bushen Chengyun granule(BCG)is an empirical treatment for female infertility(FI)caused by low endometrial receptivity(LER)involving a poorly understood mechanism.In this study,network pharmacology was used to explore the potential therapeutic mechanism of BCG on FI caused by LER.Methods:The corresponding herb targets were obtained by conducting a search in the Traditional Chinese Medicine Systems Pharmacology Database and PubMed-reported literature.Disease targets were obtained from the following databases:Comparative Toxicogenomics Database,Human Phenotype Ontology,and Therapeutic Target Database.Treatments for LER using BCG have used target matching(BCG e LER target).Then,the predicted targets were uploaded to the Search Tool for the Retrieval of Interacting Genes/Proteins database for gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses.Furthermore,triptorelin acetate for injection t menotrophin t chorionic gonadotropin for injection were used to establish a mouse model of blastocyst implantation disorder and to evaluate the in vivo effect of BCG on blastocyst implantation.Results:Overall,156 bioactive chemical components and 1092 targets of BCG were identified.The results indicated that 482 biological processes(FDR<0.01)and 15 pathways(FDR<0.01)related to BCG participated in the complex treatment effects and were associated with the endocrine system,inflammatory responses,metabolism,apoptosis,ovulatory performance,and angiogenesis.Moreover,16 hub nodes of BCG including estrogen receptor(ESR1),estrogen receptor beta(ESR2),progesterone receptor,et al,were recognized as potential treatment targets and might help clarify the underlying therapeutic mechanisms of BCG for female infertility.BCG significantly increased the protein expressions of estrogen receptors and progesterone receptors.Conclusions:These findings reveal the potential therapeutic mechanism of BCG for female infertility involves low endometrial receptivity,which should be evaluated further.

Study on the mechanism of Wenyang Jieyu Granules in the treatment of depression based on network pharmacology

Objective:To explore the potential antidepressant mechanism of Wenyang Jieyu Granules with multiple targets and components based on network pharmacology,and to verify the cAMP pathway and animal experimental results.Methods:The active components of Aconitum,Guizhi,Glycyrrhiza uralensis,Wumei,Jujube and Ginger in Wenyang Jieyu Granules were searched and screened by TCM System Pharmacology Analysis Platform(TCMSP).Using the Swiss database to predict its target,and then through the UniProt database to query the target corresponding genes;with the use of Gene Cards、OMIM database query depression related target Point and gene mapping to get the potential target of Wenyang Jieyu granule antidepressant.using Cytoscape,String software to plot the"wenyangjieyu granule-active component-potential action target-depression"network and"protein interaction network",and using David database for GO functional enrichment analysis and KEGG pathway enrichment analysis to predict the action mechanism of wenyangjieyu granule in the treatment of depression and verify it with some previous relevant experimental results.Autodock software was used to perform molecular docking experiments on selected components and targets.Results:The topology analysis is effective there are 47 core components of depression,and 40 potential key targets are SLC6A2,SHBG,ACHE,NR3C1,ESR1.they are involved in many biological processes,such as neurotransmitter receptor conduction,ammonium ion binding,postsynaptic neurotransmitter receptor activity,G protein-coupled neurotransmitter receptor activity.they play an antidepressant effect through brain tissue nerve activity ligand-receptor interaction signaling pathway,cAMP,PI3K-Akt,MAPK signaling pathway,ErbB signaling pathway,etc.At the same time,the results of molecular docking show that the core components have good binding force to the key targets.Conclusion:Wenyang Jieyu granule.The active components of the anti-depressant effect are mediated by multiple signaling pathways and multiple targets.

Potential active compounds and mechanisms of Shen-Qi-Yi-Chang granule for the treatment of colorectal cancer:an analysis of network pharmacology and molecular docking

Background:In recent years,herbal formulations have assumed an influential part in preventing and treating tumors.Shenqi Yichang granules(SQYCG)have proven effective in the adjuvant treatment of colorectal cancer(CRC),but their mechanism has not been elucidated.This study aimed to explore the potential active compounds and mechanisms of SQYCG in the treatment of CRC using network pharmacology and molecular docking.Methods:The active compounds and targets of SQYCG and the CRC genes were found using the Traditional Chinese Medicine Systems Pharmacology,DrugBank,and DisGeNET databases.The intersected targets of disease genes and drug targets were depicted using a Venn diagram.The protein-protein interaction(PPI)network of these targets was obtained by String platform and visualized using Cytoscape.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were carried using the DAVID database to obtain the core molecular mechanism of SQYCG in CRC treatment.Molecular docking techniques were used to validate the results.Results:A total of 63 compounds and 245 targets were obtained from the herbal prescription after the screening,of which 122 targets crossed with CRC genes.PPI showed that the core regulatory targets include MAPK1,TNF,TP53,JUN,RELA,MAPK14,and MAPK 8.The GO analysis indicated regulation of drug response,apoptotic process,response to hypoxia,angiogenesis,and response to lipopolysaccharide.KEGG pathway enrichment analysis mainly involves TNF,T cell receptor,Toll-like receptor,PI3K-Akt,and MAPK signal pathway.Conclusion:Through network pharmacology,we havedemonstrated that SQYCG has multiple targets,components,and pathways in treating CRC,with anti inflammation and inhibition of cell proliferation being critical components of its mechanism.

Exploring the mechanism of Pinggan Yishen granules in the treatment of hypertension and insomnia through network pharmacology and molecular docking

Background:Hypertension is closely related to insomnia.Pinggan Yishen(PGYS)Granule is an effective compound medicine for treating hypertension and insomnia.In this study,we aimed to systematically explain the potential mechanism of PGYS granules in the treatment of hypertension and insomnia using network pharmacology and molecular docking techniques.Methods:Potential targets accounted for hypertension and insomnia were obtained from OMIM,GeneCards,and DrugBank databases.We used the Batman-TCM database to query natural compounds and potential targets associated with PGYS granules.According to the localization results of PGYS granules and potential target genes of disease,the protein-protein interaction network was constructed.The tissue and subcellular distribution information for key proteins are visualized.Used KOBAS3.0 to enrich KEGG pathways and GO biological processes.Molecular docking was used to verify relationships between core compounds and proteins.Results:The comorbid mechanism of hypertensive insomnia is mainly related to disorders of neurotransmitter regulation,imbalance of the renin-angiotensin-aldosterone system(RAAS),abnormal metabolism including cytokine secretion and lipid metabolism.The potential therapeutic mechanisms of PGYS granules include the regulation of neurotransmitters,lipid metabolism and inflammatory response.CACNA1C,adrenergic receptors,cytochrome P450 family and muscarinic cholinergic receptors may be the key targets of PGYS granules.Conclusion:Hypertension and insomnia are related in mechanisms.PGYS granules may play a therapeutic role in hypertension and insomnia by regulating neurotransmitters,lipid metabolism and inflammatory response.

Mechanism of Bushen Tongluo granule on osteoarthritis based on network pharmacology and molecular docking technology

Objective:In this study,we used network pharmacology and molecular docking technology to analyze the mechanism of Bushen Tongluo granule in the treatment of osteoarthritis.Methods:The main active components and corresponding targets of Bushen Tongluo granule were screened from thetraditional Chinese medicine systems pharmacology database.The targets related to osteoarthritis were collected from the Online Mendelian Inheritance in Man,Therapeutic Target database,GeneCards,Pharmacogenomics Knowledgebases and Drugbank databases.Cytoscape3.9.0 software was used to construct the action network diagram of“Bushen Tongluo Granule-Active Component-Target”.Builded a protein-protein interaction network from the STRING database.The Bioconductor platform and R language 4.0.3 were used for Gene Ontologyfunction andKyoto Encyclopedia of Genes and Genomespathway enrichment analysis.Then,selected the pathway most associated with osteoarthritis for specific analysis.Finally,the core genes were screened and verified by molecular docking using AutoDockTools software.Results:71 principal components of Bushen Tongluo granule and 183 potential therapeu-tic targets for osteoarthritis were obtained.Twenty-eight key targets of Bushen Tongluo granulein the treatment of osteoarthritis are enriched in 158 pathways.Among them,the tumor necrosis factor signaling pathway,interleukin-17 signaling pathway,Toll-like receptor signaling pathway,T helper cell 17 cell differentiation and hypoxia-inducible factor-1signaling pathway involved in key targets are closely related to osteoarthritis.The relevant vital targets were involved in the regulation of DNA transcription factor activity,the response to chemical stress,the response to reactive oxygen species,the response to oxidative stress,the proliferation of muscle cells,the proliferation of epithelial cells and the biological processes such as responses to lipopolysaccharides,responses to molecules of bacterial origin.Molecular docking showed that protein kinase B 1-β-sitosterol,the tumor necrosis factor-naringenin,interleukin-6-luteolin,mitogen-activated protein kinase-quercetin,vascular endothelial growth factor A-quercetin and prostaglandin-endoperoxide synthase 2-luteolin have strong docking activities.Conclusions:Bushen Tongluo granule can reduce the inflammatory response and inhibit articular cartilage angiogenesis in the treatment of osteoarthritis,which may be achieved by regulating the inflammatory signaling pathway and hypoxia-inducible factor-1 signaling pathway.

Network Pharmacology-based Analysis on Determining the Mechanisms of Yishen Qutong Granules(益肾祛痛颗粒)in Alleviating Cancer-related Fatigue

Cancer-related fatigue(CRF)is associated with cancer-related anemia(CRA).As the common comorbidities of cancer,both of them can seriously affect the quality of patient life.Yishen Qutong Granules(益肾祛痛颗粒,YSQTG)have achieved good curative effects in the treatment of CRA.However,the mechanism of whether it can alleviate CRF needs further confirmation.We used network pharmacology and molecular docking to investigate the molecular mechanism and the effective compounds of the prescription.Through the analysis and research in this paper,we obtained 76 effective compounds and 76 drug-disease intersection targets to construct a network,indicating that quercetin,luteolin,baicalein,β-sitosterol and stigmasterol were possibly the most important compounds in YSQTG.The key targets of YSQTG for CRF were mainly enriched in IL-17 and TNF pathways.816 GO entries and 113 pathways were obtained by GO and KEGG enrichment,respectively,which proved that YSQTG might have a comprehensive therapeutic effect on CRF mainly through regulating IL-17,TNF,MAPK,NF-κB and chemokines,as well as cholinergic synapse and 5-HT synapse pathways.The results of molecular docking showed thatβ-sitosterol and stigmasterol could form PI-Alkyl or Alkyl hydrophobic interactions with CXCL8 and ESR1 at residues LEU25,ARG26,PHE65,ALA69 and LEU346,ALA350,LEU391,PHE404,LEU525,VAL533,respectively.In conclusion,the therapeutic effect of YSQTG on CRF is based on the comprehensive pharmacological effect of multicomponent,multitarget,and multichannel pathways.This study provides a theoretical basis for further experimental research.

Investigating the Potential Mechanism of Huazhi Rougan Granules Against Alcoholic Liver Disease by Network Pharmacology and Experiment

Objective: Huazhi Rougan granules(HRGs) are a promising drug to improve alcoholic liver disease(ALD), but its mechanism remains unclear. Materials and Methods: Network pharmacology, molecular docking, and animal experiments were used to reveal the potential mechanism of HRG against ALD. Results: A total of 245 potential targets of HRG against ALD were obtained. Functional enrichment analysis suggests that HRG could synergistically regulate various biological pathways to exert therapeutic effects on ALD. Molecular docking showed that the key targets exhibited the good binding ability with the key compounds. The in vivo experiments showed that HRG can effectively alleviate pathological changes in liver tissue, improve blood lipid levels, antioxidant stress ability, and liver function, and reduce the release of inflammatory cytokines in the liver. Conclusions: This study reflects that HRG is an effective strategy for treating ALD, providing a basis for revealing the prevention and treatment mechanisms of ALD.

Mechanism of Mingjing Granules in Treating Wet Age-Related Macular Degeneration Based on Network Pharmacology and Experimental Verification

Objective:To analyze the potential mechanism of Mingjing granules in the treatment of wet age-related macular degeneration(wAMD)based on the research methods of network pharmacology and molecular docking approach and to provide a new reference for the currently limited treatment of wAMD.Materials and Methods:We searched TCMSP,GeneCards,OMIM,PharmGkb,TTD,and DrugBank database to screen the main active ingredients of Mingjing granules and their therapeutic targets of wAMD.The network of active components and targets was constructed using Cytoscape3.6.1 software,which was also used for the topological analysis of target genes.The network of Protein-Protein Interactions(PPI)was mapped using the String platform.We also used R language to do the Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway for additional analysis.Molecular docking studies were finished by Chemoffice,Autodock,and Pymol.Finally,the efficacy of the Mingjing granules was examined in animal experiments,in which we used enzyme-linked immunosorbent assay to the contents of vascular endothelial growth factor(VEGF)and matrix metalloproteinase-9(MMP-9)levels in peripheral blood.Results:Active compounds,including quercetin,lignocaine,and kaempferol,were found.PPI network analysis showed that tumor necrosis factor(TNF),MMP-9,epidermal growth factor(EGF),prostaglandin-endoperoxide synthase 2(PTGS2),and caspase-3(CASP3)were related to both Mingjing granules and wAMD.GO and KEGG pathway analysis showed that these targets were mainly involving lipids and atherosclerosis,TNF,and interleukin-17(IL-17)signaling pathways.Docking studies suggested that quercetin and luteolin can fit in the binding pocket of four target proteins(CASP3,EGF,PTGS2,and TNF).In the vivo experiment,the Mingjing granules were found to be effective on the expression of VEGF and MMP-9 in peripheral blood.Conclusions:This study initially reveals the multi-constituent,multi-target,and multi-pathway mechanism of action of Mingjing granules in the treatment of wAMD and implies the inhibition of choroidal neovascularization may be related to the expression of VEGF and MMP-9.

Efficacy of Ganshuang granules(肝爽颗粒)on non-alcoholic fatty liver and underlying mechanism:a network pharmacology and experimental verification

OBJECTIVE:To investigate the potential pharmacological mechanisms of Ganshuang granules(肝爽颗粒,GSG)in treating non-alcoholic fatty liver(NAFLD).METHODS:All the active components and targets of GSG were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.Protein-Protein interaction network,Kyoto Encyclopedia of Genes and Genomes and Gene Ontology function annotation of common targets were analyzed to predict the mechanisms of action of GSG in the treatment of NAFLD.Then,the mouse models of NAFLD were constructed in a diet-induced manner and treated with GSG.The levels of interleukin 6(IL-6),tumor necrosis factor-alpha(TNF-α)and phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT)pathway-related proteins in the liver of mice in each group were measured by enzyme linked immunosorbent assay and Western blot,respectively.RESULTS:Network pharmacology revealed a total of 159 potential targets of GSG for the treatment of NAFLD.Functional enrichment analysis indicated that the PI3K/AKT signaling pathway may be involved during GSG treatment of NAFLD.Further experiments showed that the significantly decreased alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase,total cholesterol,triglyceride and low-density lipoprotein cholesterol levels in NAFLD model mice serum after GSG treatment,as well as the expression levels of IL-6 and TNF-αin the liver.Furthermore,drug intervention increased the protein expression levels of phosphorylated-PI3K(P-PI3K)and P-AKT in the liver of the model group mice,and decreased the protein expression level of sterol regulatory element-binding protein 1.CONCLUSION:We found that GSG is effective in treating NAFLD and the potential therapeutic targets may be involved in PI3K/AKT signaling pathway.

Network Pharmacology and in vitro Experimental Veriflcation on Intervention of Quercetin, Present in Chinese Medicine Yishen Qutong Granules, on Esophageal Cancer

Objective: To explore the potential mechanism of Yishen Qutong Granules(YSQTG) for the treatment of esophageal cancer using network pharmacology and experimental research. Methods: The effective components and molecular mechanism of YSQTG in treating esophageal cancer were expounded based on network pharmacology and molecular docking. The key compound was identified by high-performance liquid chromatography and mass spectrometry(HPLC-MS) to verify the malignant phenotype of the key compounds in the treatment of esophageal cancer. Then, the interaction proteins of key compounds were screened by pull-down assay combined with mass spectrometry. RNA-seq was used to screen the differential genes in the treatment of esophageal cancer by key compounds, and the potential mechanism of key compounds on the main therapeutic targets was verified. Results: Totally 76 effective compounds of YSQTG were found, as well as 309 related targets, and 102 drug and disease interaction targets. The drug-compound-target network of YSQTG was constructed, suggesting that quercetin, luteolin, wogonin, kaempferol and baicalein may be the most important compounds, while quercetin had higher degree value and degree centrality, which might be the key compound in YSQTG. The HPLC-MS results also showed the stable presence of quercetin in YSQTG. By establishing a protein interaction network, the main therapeutic targets of YSQTG in treating esophageal cancer were Jun proto-oncogene, interleukin-6, tumor necrosis factor, and RELA proto-oncogene. The results of cell function experiments in vitro showed that quercetin could inhibit proliferation, invasion, and clonal formation of esophageal carcinoma cells. Quercetin mainly affected the biological processes of esophageal cancer cells, such as proliferation, cell cycle, and cell metastasis. A total of 357 quercetin interacting proteins were screened, and 531 genes were significantly changed. Further pathway enrichment analysis showed that quercetin mainly affects the metabolic pathway, MAPK signaling pathway, and nuclear factor kappa B(NF-κB) signaling pathway, etc. Quercetin, the key compound of YSQTG, had stronger binding activity by molecular docking. Pull-down assay confirmed that NF-κB was a quercetin-specific interaction protein, and quercetin could significantly reduce the protein level of NF-κB, the main therapeutic target. Conclusion: YSQTG can be multi-component, multi-target, multi-channel treatment of esophageal cancer, it is a potential drug for the treatment of esophageal cancer.

Molecular mechanism of Mimenghua Granules in treating dry eye

Objective:To explore the molecular mechanism of Mimenghua Granules in treating dry eye based on network pharmacology and bioinformatics methods.Methods:Screening and prediction of possible blood-inducing active ingredients and action target of Mimenghua Granules through Traditional Chinese Medicine Systems Pharmacology database and analysis platform;mining dry eye-related diseases through disease gene database,gene target;use the functional protein combined network database STRING to draw the component-target and disease-target PPI networks,and extract the intersection of these two networks;use DAVID database analysis to screen key targets and analyze the mechanism of action.Results:A total of 593 active ingredients related to Mimenghua Granules were retrieved from the Traditional Chinese Medicine Systems Pharmacology database,and 59 blood active ingredients were obtained by screening based on pharmacokinetic parameters,and 680 targets related to these ingredients were retrieved;from disease genes,the database searches for 47 genes directly related to dry eye;3 key genes(ICAM1,IFNG,and IL-6)were obtained after the intersection of the component target and disease target PPI network;these genes are mainly involved in natural killer cell-mediated cytotoxicity,Jak-STAT signaling pathway,and Cytokine-cytokine receptor interaction.Conclusion:The mechanism of Mimenghua Granules in treating dry eye is related to the interaction of cytotoxic pathway,Jak-STAT signal pathway,and cytokines.The key gene targets are ICAM1,IFNG,and IL-6.