基于网络药理学和实验验证探讨加味丹栀逍遥散治疗抑郁症的作用机制OACSTPCD

Objective:To use network pharmacology methods to explore the mechanism of action of modified Danzhi Xiaoyao Powder in the treatment of depression,and to verify the potential targets of modified Danzhi Xiaoyao Powder in the treatment of depression through molecular docking and animal experiments.Method:Based on the Traditional Chinese Medicine System Pharmacology Database and A-nalysis Platform(TCMSP)and the Encyclopedia of Traditional Chinese Medicine(ETCM),the chemical components and target of ac-tion of modified Danzhi Xiaoyao Powder were screened.DisGeNet and the Online Mendelian Inheritance Inman(OMIM)database are used to search for depression related targets,and use STRING to expand secondary targets to obtain all targets of depression.Intersect the targets related to the active ingredients of modified Danzhi Xiaoyao Powder with the targets of depression to obtain the targets related to the treatment of depression with modified Danzhi Xiaoyao Powder.Network Cytoscape software was used to a protein-protein interac-tion(PPI).Perform gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signal pathway enrichment analysis on potential targets by using DAVID.Molecular docking of core targets and components through AutoDock.48 SPF grade male C57BL/6 mice were randomly divided into a normal group of 8 mice and a modeling group of 40 mice.A chronic unpredicta-ble mild stress(CUMS)model was established in the modeling group of mice.After 6 weeks,the modeling group of mice was divided into a CUMS model group,a fluoxetine group(20 mg·kg-1),and a modified Danzhi Xiaoyao powder low(15.9 g·mL-1),medium(31.8 g·mL-1),and high(47.7 g·mL-1)dose group,with 8 mice in each group.The corresponding drugs were administered by gavage at a volume of 10 mL·kg-1.The normal group and the model group were compared.Administer equal volume of physiological saline by gavage once a day for 28 days.The behavioral ability of mice was evaluated by using sucrose preference experiments,tail sus-pension experiments and open field experiments.HE staining was used to observe the pathological changes in the hippocampus of the mouse brain;Observation of changes in newly generated neurons in the hippocampus using DCX staining;Western Blot was used to de-tect the expression levels of PI3K,p-PI3K,AKT,p-AKT,and BDNF proteins.Results:Network pharmacology analysis showed that there were 164 active ingredients in modified Danzhi Xiaoyao Powder,corresponding to 332 targets;1 432 depression related targets;By taking the intersection of the targets related to modified Danzhi Xiaoyao Powder and those related to depression,147 potential targets for the treatment of depression with modified Danzhi Xiaoyao Powder were identified;PPI network analysis identified 9 core targets;KEGG enrichment analysis mainly involves cell apoptosis,phosphatide3-kinase(PI3K)-protein kinase B(PKB,also known as AKT)sig-naling pathway,etc.The molecular docking results indicate that kaempferol,naringenin,and quercetin have the strongest binding ability with AKT.The animal experiment results showed that compared with the normal group,the CUMS group mice showed a significant de-crease in sugar water preference rate,tail suspension immobility time,and p-AKT and BDNF protein expression levels(P＜0.01);Compared with the CUMS group,the sugar water preference rate,tail suspension immobility time,and p-AKT and BDNF protein ex-pression levels of mice in the modified Danzhi Xiaoyao powder group were significantly increased(P＜0.05).HE staining showed that the neuronal structure of the CA3 area in the hippocampus of CUMS group mice was loose,the number of cells decreased,and the ar-rangement was irregular;The neuronal structure of the CA3 region in mice treated with modified Danzhi Xiaoyao powder was more com-plete,and the number of cells was close to that of the normal group.Immuno fluorescence showed a significant decrease in the number of DCX positive cells in the DG area of the hippocampus of CUMS mice;The number of DCX positive cells in the DG area of the hippo-campus of mice treated with modified Danzhi Xiaoyao powder significantly increased.Conclusion:Modified Danzhi Xiaoyao Powder treat-ment for depression has the characteristics of multiple components,multiple targets,and multiple pathways,and its mechanism of action may be related to promoting cell proliferation and affecting neuronal regeneration.