Study On Fructus Corni And Cortex Moutan Against Liver Injury Based On Network Pharmacology And Bioinformatics Research On Disease Targets

Objective:By using databases and platforms of network pharmacology,the potential active components and targets of Fructus Corni and Cortex Moutan against liver injury were screened,and the possible functional routes were analyzed.The potential biomarkers of nonalcoholic steatohepatitis and the possible small molecule drugs were screened by using bioinformatics technology.Methods:TCMSP database was used to search and screen the active components of Fructus Corni and Cortex Moutan.Then we collected the SMILE structure formula and SDF structure file of each compound in PubChem,and predicted the composition targets by using SwissTargetPrediction,Drugbank and SEA database respectively.Then,we searched for keywords such as "liver disease","liver injury","liver injury","liver" and "hepatic" in the disease database(TTD,GAD and DisGeNET)to collect the targets of liver injury,and removed the duplicate parts after combination.By intersecting component target set and disease target set,the corresponding disease target of each component was obtained.Finally,the enrichment of GO and KEGG were analyzed by DAVID platform.All the above operations were under human database.The data of compositions,targets and pathes were imported into the software of Cytoscape to build the network,and the core compositions and targets in the network were selected according to the degrees.In addition,String database was used to build the relationship between the targets,and we selected the Top9 central targets in PPI network.GEO database and ROC curve were used to evaluate the correlation between the central targets and liver injury in the process of nonalcoholic steatohepatitis.To search the data set of nonalcoholic steatohepatitis in GEO database,clinical samples and healthy control group were required.After downloading the original data set through the"Geoquery" package in the R language software,we replaced the missing value with 0 and analyzed the sample grouping quality with PCA.Differential genes of the data sets were screened by plug-in GE02R,and the standard were |log2(FC)|>1 and adj.P<0.05.Visualization was implemented with R package "ggplot2".Through RRA algorithm,the differential genes of each data set were integrated.The standard were |log2(FC)|>1 and P<0.05 which was corrected by Bonferroni.After integration,the differential genes were enriched and analyzed by DAVID platform,and visualized by R package "GOplot".The relationships between genes were constructed by String database,and the network was constructed by Cytoscape.The Top 10 core targets in the network were selected by the plug-in cytohubba and MCC algorithm,and cluster analysis was carried out by using the plug-in clusterONE.In CTD database,the core targets and the ways of enrichment analysis were verified.Finally,the cMAP database and the affymetrix platform were used to achieve relevant small molecule drugs matched with differential genes.Results:The active components of Fructus Corni and Cortex Moutan were 20 and 11,respectively.There were 776 and 625 component targets,respectively.A total of 667 disease targets were sorted out.After the intersection,52 anti-liver injury targets of Fructus Corni and 69 of Cortex Moutan were obtained,and totally 78 targets were obtained after combination and deduplication.In the enrichment analysis of GO and KEGG,results of GO-BP included steroid metabolic process,xenobiotic metabolic process,steroid hormone mediated signaling pathway,etc.GO-CC included organelle membrane,extracellular space,membrane raft,etc.GO-MF included RNA polymerase II transcription factor activity,steroid hormone receptor activity,enzyme binding,etc.KEGG showed that there were HIF-1 signaling pathway,TNF signaling pathway,osteoclast differentiation,bile secretion,insulin resistance and other pathways.The core components of the network were quercetin,kaempferol,?-sitosterol,sitosterol,stigmasterol,etc.The core targets were AKT1,PIK3CG,PTPN1,MAPK8,PPARA,ABCB1,TNF,JUN,EGFR,CYP2C19,etc.The results of ROC curve evaluation showed that the areas under the curve of VEGFA,ESR1 and MAPK8 were>0.70?0.90,and the expressions were significantly different(P<0.05).GSE17470,GSE24807,GSE37031 and GSE89632 were included in the study.The results of principal component analysis showed that each data set had a good differentiation between NASH group and control group.The RRA algorithm finally identified 59 differential genes,including 39 up-regulated genes and 20 down-regulated genes.The results of GO analysis showed that GO-BP had cellular response to hormone stimulus,female pregnancy,inflammatory response,etc.GO-CC had extracellular space,extracellular region and endoplasmic reticulum,etc.GO-MF had transcription factor activity,receptor binding,insulin-like growth factor II binding,etc.The Top 10 core targets were FOS,ATF3,KLF4,CDKN1 A,GDF15,KLF6,FOSB5 GADD45B,CRP and ZFP36.The possible small molecular drugs were doxorubicin,tanespimycin,fulvestrant,sitosterol and so on.Conclusion:1.Fructus Corni and Cortex Moutan have multi-component and multi-target effects on liver injury,and the potential active components are mainly flavonoids and sterols,which may become the candidate components of liver protection drugs.The potential targets may be the targets of Fructus Corni and Cortex Moutan on liver injury,which still need to be studied.2.Based on the comprehensive bioinformatics analysis,10 core genes related to NASH are screened out.The study of these genes will help us to further understand the progress and mechanism of NAFLD and NASH.The selected small molecule drugs may become the candidate drugs for the treatment of diseases.In addition,these core genes may be potential biomarkers and therapeutic targets for NAFLD or NASH,however further preclinical and clinical trials are still needed.