Exploring the action mechanism of anti-depression of Zhishi Zhizichi Soup based on network pharmacology and experimental validation
Objective To predict the action target of anti-depression and signalling pathway of Zhishi Zhizichi Soup by network pharmacology and to experimentally validate it.Methods The active ingredients of Zhishi Zhizichi Soup were screened and their action targets were predicted using the Systems Pharmacology database andanalysisplatform thepotential disease targets of depression were retrieved through OMIM and GeneCards,and the intersecting targets of the ingr-edients and the diseases were selected as the protein target libraries,and STRING was used to obtain the network of the intersecting target-protein interactions,and the data were analysed through the Cytoscape software for the Visualisation analysis was performed to construct the active ingredient-predicted target-signalling pathway network.The key targets of Zhishi Zhizichi Soup were analysed by proteinprotein interaction and animal experiments were performed.Meanwhile,gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed using the DAVID database to comprehensively evaluate the antidepressant mechanism of action of Zhishi Zhizichi Soup.After 30 SD rats were acclimatised and fed for one week,they were randomly divided into six in the normal group and 24 in the modelling group.Except for the normal group,the experimental rats were subjected to chronic unpredictable mild stress,and after confirming the success of modelling,the modelling group was divided into four groups according to the random number table method,the high-dose group(13.32 g/kg Zhishi Zhizichi Soup),the medium-dose group(6.66 g/kg Zhishi Zhizichi Soup),the low-dose group(3.33 g/kg Zhishi Zhizichi Soup),and model group,with six animals in each group,on which animal behavioural and Western blotexperiments were performed.Results Thirty-eight effective active ingredients of Zhishi Zhizichi Soup were screened,and 52 potential targets of Zhishi Zhizichi Soup for the treatment of depression were predicted,and the key targets were screened to include ERK,CREB,and BDNF as the validation pathways.The results of GO and KEGG analyses showed that the treatment of depression by Zhishi Zhizichi Soup involved several signalling pathways such as FoxO signalling pathway,MAPK signalling pathway,prolactin signalling pathway,oestrogen signalling pathway,ErbB signalling pathway,GnRH signalling pathway,sphingolipid signalling pathway and so on.The results of the experimental animal behavioural experiments showed that compared with the normal group,the sugar-water preference rate of the model group was decreased,the swimming immobility time was increased,and the horizontal and vertical scores of the absenteeism experiment were reduced(P＜0.01).Compared with the model group,sugar-water preference was increased in the low-dose and medium-dose groups(P＜0.05);compared with the model group,swimming immobility time was decreased in the medium-dose and high-dose groups(P＜0.05);and compared with the model group,the vertical scores of the low-dose,medium-dose,and high-dose groups were increased(P＜0.05);compared with the model group,the level scores of both low-dose and medium-dose groups were increased(P＜0.01).The results of Western blot showed that the protein expression levels of p-ERK1/2,p-CREB,and BDNF in the hippocampus of rats in the model group were reduced compared with those in the normal group(P＜0.01).Compared with the model group,the p-CREB protein expression level was increased in the low-dose,medium-dose,and high-dose groups(P＜0.05);compared with the model group,the BDNF protein expression level was increased in the low-dose and medium-dose groups(P＜0.05).Conclusion Zhishi Zhizichi Soup can improve depressive behaviour in rats and has therapeutic effects on depression.The mechanism by which it exerts antidepressant effects is related to ERK,CREB,and BDNF.
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