Based On Network Pharmacology Method To Explore The Mechanism Of Erxian Decoction In Polycystic Ovary Syndrome

ObjectiveThe Erxian decoction(EXD)group was studied by network pharmacology method to construct a database of chemical constituents of traditional Chinese medicine,confirming the active compounds and targets in Erxian Decoction,and using bioinformatics methods to mine polycystic ovary syndrome(Polycystic ovary syndrome(PCOS)related pathogenic gene,constructing EXD-PCOS protein interaction network,analyzing the key targets of EXD treatment of PCOS and bioinformatics analysis of its biological function,exploring the potential mechanism of Erxian decoction in the treatment of PCOS,and It carries out experimental verification,provides theoretical support for clinical rational drug use,and provides experimental basis and basis for further experimental research.Method1.Establishment the database of EXDThe chemical constituents of Traditional Chinese Medicine in Erxian Tang were retrieved from TCMID,TCMSP and TCM@taiwan.The chemical composition of Erxian Decoction was constructed and the active compounds were screened according to OB and DL.Target prediction was performed using random forests and support vectors.All compounds and targets were uniformly named using Pubchem and Uniprot for subsequent studies.2.Network establishedA PCOS-related disease database was constructed based on the NCBI-GENE and OMIM database to retrieve PCOS-related genes.Through the protein interaction website STITCH,the related gene protein interactions were constructed to form disease genes interaction network.Compound-target,target protein interaction network,and disease-related gene interaction network were visualized using Cytoscape software.Analyze network attributes and topological parameters to provide an in-depth analysis of EXD components and diseases.3.Bioinformatics analysis of the key targetsThe EXD interaction target interaction network was mapped into the disease-related gene protein interaction network,and the key targets of drug treatment PCOS were confirmed,then analysis the GO and KEGG bioinformatics analysis by was the ClueGO plug-in of cytoscape to explore the biological mechanism of treatment of PCOS.4.ExperimentSixty SD rats,21 days old.Randomly divided into 2 groups,10 in the control group and 50 in the model group.The amount of DHEA in the model group was used to induced PCOS rats,continuous injection for 20 days;detection of ovulation and serum FSH,LH test model,selected successful rats were randomly divided for three groups.Model group: Gave water stomach and saline intravenous injection;Treatment group: Treated with Erxian Decoction and the saline was injected into the tail vein;Inhibitor group: Erxian Decoction.+LY294002 was injected into the tail vein of rats.After 3 weeks,the recovery of estrous cycle was observed.The changes of serum T,E2,FSH,LH,FINS and LH/FSH were detected by ELISA.The morphology of ovary was analyzed by HE staining.Immunohistochemistry,Western Blot was used to detect the expression of p-PI3 K,p-AKT and GSK3? protein in rat ovarian tissue.Result1.A total of 981 Erxian decoction compounds were obtained from the Chinese medicine database.According to analyze and arrange,70 active compounds were selected according to OB and DL screening conditions,corresponding to 247 targets;262 PCOS-related genes were searched by NCBI-GENE and OMIM database.Construct a compound-target network,a target interaction network,and a related gene action network,respectively.2.Mapping the drug target network to the disease gene interaction network can obtain 50 co-targets.These co-targets may be the key targets for the treatment of PCOS by Erxian decoction,GO analysis and KEGG biological pathway for 50 key targets.Analysis,these key targets are mainly involved in the apoptotic metabolic pathway,regulation of reactive oxygen metabolism,protein kinase B signaling pathway,regulation of sequence-specific DNA-binding transcription factor activity;mainly involved in cellular components of platelet alpha particles,RNA polymerase II transcription factors,Plasma membrane raft;the main molecular functions involved include cytokine activity,protein phosphatase binding,and activation of steroid hormone receptors.Enrichment of key targets for KEGG biological pathways revealed that these key targets are involved in biological pathways including: AGE-RAGE signaling pathway,fluid shear stress and atherosclerosis,PI3K-Akt signaling pathway,HIF-1 signaling pathway,alcoholic fatty liver(NAFLD)and MAPK signaling pathway and others total 15 biological pathways.3.Compared with the normal control group,the serum LH and FSH levels in the PCOS model group were increased,LH/FSH>2,and the estrous cycle was disordered,and the modeling was successful.Compared with the model group,the rats in the EXD treatment group established regular estrous cycle,serum FSH increased and serum T,LH and FINS metabolic indexes were significantly decreased,LH/FSH<2,the treatment group was superior to the inhibitor group;ovarian tissue HE Staining showed that the follicles in the model group were severely dilated,the radiation crown disappeared,and there was no dominant follicle.The treatment group and the inhibitors had a mild and moderate expansion.The corpus luteum and atresia follicles,oocytes and radioactive crown were observed.The treatment group was superior to the inhibitor group.EXD can effectively improve serum hormone levels in PCOS rats,and has a certain therapeutic effect on PCOS.PI3K/AKT pathway blocker(LY294002)can block the effect of EXD on the reproductive hormone disorder and IR of PCOS.ConclusionThis study integrates multiple databases to construct EXD Chinese medicine database using network pharmacology method,screens related genes of PCOS through data mining,and uses network pharmacology to create compound-target network,protein-protein interaction network,and drugdisease related gene protein interaction network.The basic properties and topological parameters of the network were analyzed,and the mechanism of EXD was explored.The drug-target network was mapped into the gene protein interaction network tool.The mechanism of EXD treatment of PCOS was analyzed and verified by experiments.This method provides a theoretical basis for clinical rational drug use and experimental research,and provides new ideas for Chinese herbal medicine research.