Study On The Target And Animal Verification Of Chaihu Shugan Decoction On Depressed Rats Based On Network Pharmacology

Objective: To screen out the key targets of the ancient and modern famous prescription "Chaihu Shugan Decoction" and depression,and reveal the mechanism of action of Chaihu Shugan Decoction in treating depression by using the method of network pharmacology.Then through molecular docking technology,so as to reveal the molecular mechanism of Chaihu Shugan Decoction in treating depression from the molecular level.Finally,the selected core targets were verified by animal experiments,which provided ideas and certain objective basis for the clinical application of Chaihu Shugan Decoction and providing experimental basis for the follow up study of Chaihu Shugan Decoction in the treatment of de-pression.Method: Chaihu Shugan Decoction was composed of Chaihu,Zhike,Shaoyao,Gancao,Xiangfu,Chuanxiong and Chenpi.The effective active components of the above seven drugs were searched on the platform of TCMSP,and the corresponding action targets were screened out.The gene symbol and Uniprot number of the drug action target are obtained on the Uniprot database,and they are standardized.The visualization diagram of the effective active component-action target network was drawn through the Cytascape software,and the core effective,components of Chaihu Shugan Decoction were screened out.Disease targets of depression were found in Gene Cards,OMIM,TTD,Pharm GK,Dis Ge NET and Drug Bank databases,and standardized treatment of disease targets was conducted through Uniprot database.Search the disease targets of depression in Gene Cards,OMIM,TTD,Pharm GK,Dis Ge NET and Drug Bank databases,and standardize the disease targets through Uniprot database.Venny online mapping software is used to draw the Waynediagram of drug action target and depression disease target,and the common target of drug and disease is obtained.The common targets of drugs and diseases were put into the STRING database,and the protein-protein interaction network was constructed to screen out the key targets and core target groups.The core target was imported into Metascape database,and the GO function and KEGG enrichment pathway of the core target were analyzed,and the network diagram of core effective components-target-pathway was drawn by Cytoscape software,from which the core effective components with high correlation were screened out for molecular docking with the target,and the docking results were analyzed from the molecular level.In animal experiment,rats were divided into control group,depression group and Chaihu Shugan Decoction group.Rats’ depression model was established by lipopolysaccharide,and each group was given corresponding intervention methods.The behavior of rats in each group was tested by open field experiment and sugar water preference test,and the protein expression level and relative mRNA expression of the core target were detected and verified by WB and fluorescence PCR.Results: 1.After screening,139 active ingredients and 452 potential drug targets of Chaihu Shugan Decoction were obtained,and 1070 disease targets of depression were screened,with 92 common targets.2.GO function and KEGG enrichment pathway analysis showed that core targets were involved in 714 biological processes,18 cell components,50 molecular functions,and143 signaling pathways related to depression,such as inflammation,immunity,cell proliferation,cell differentiation,and metabolism.3.The results of molecular docking showed that the key target had good binding activity with the core components.4.In terms of animal experiments,(1)In terms of body mass,7 days after modeling,body mass of rats in model group was significantly reduced compared with control group,P<0.001;Compared with model group,body weight of rats in Chaihu Shugan Decoction group was increased(P<0.05).On 14 days and 21 days after modeling,the body mass of rats in the model group was significantly reduced compared with the control group,P<0.001;Compared with model group,body weight of rats in Chaihu Shugan Decoction group was significantly increased(P<0.001).(2)In the test of sugar water preference and open field experiment,compared with the control group,the volume of sugar water consumption(ml),the preference rate of sugar water(%),the total movement distance(CM),the central movement distance(%)and the central residence time(%)of rats in model group were significantly decreased,P<0.001;Compared with model group,the volume of sugar water consumption(ml),the preference rate of sugar water(%)and the movement distance in the central area(%)of rats in Chaihu Shugan Decoction group were significantly increased(P<0.001),and the total movement distance and the residence time in the central area(%)were significantly increased(P<0.01).(3)Compared with the control group,IL-6protein expression level and mRNA expression level in hippocampal tissue of model group were significantly increased(P<0.001),while AKT1,CREB1,FOS protein expression levels and mRNA expression levels were significantly decreased(P<0.001);Compared with model group,the protein expression level and mRNA expression level of IL-6 in hippocampus of rats in Chaihu Shugan Decoction group were significantly decreased(P<0.001),while the protein expression levels and mRNA expression levels of AKT1,CREB1,FOS were significantly increased(P<0.001).Conclusions:1.The treatment of depression by Chaihu Shugan Decoction is the result of multi-component,multi-target and multi-pathway.2.Through network pharmacology and molecular docking,it is found that Chaihu Shugan Decoction could be inhibit the activation of inflammatory factors,mediate neurotrophic factors,inhibit nerve cell apoptosis and protect neurons by acting on relevant targets and pathways,thus achieving the effect of anti-depression.3.The results of animal experiments showed that Chaihu Shugan Decoction could significantly restore the body weight and activity ability of depressed rats.The treatment of depression might be through down-regulating the expression level of IL-6 protein and decreasing the relative expression level of IL-6mRNA.Up-regulated expression levels of AKT1,CREB1 and FOS protein,increased r-elative expression levels of AKT1 mRNA,CREB1 mRNA and FOSmRNA,so as to improve depressive symptoms,thus affecting the occurrence and development of depression.