Abstract: Objective In this study, we aimed to use network pharmacology techniques to predict the key targets of a prescription of Ziziphi spinosae semen formula (ZSSF) compound for depression, and to verify its mechanism of action using a zebrafish model of rifampicin-induced depression. Methods The drug targets of ZSSF were retrieved from the TCMSP database, and the target names were corrected using the UniProt database. Depression-related targets were identified using the GeneCards, OMIM, and NCBI databases. Protein-protein interaction information for the shared targets was predicted using the STRING database. The collected data were then analyzed using the Metascape database to determine GO and KEGG pathway enrichment, and the result were visualized using microbiotics. Behavioral experiments and reverse-transcription quantitative PCR experiments were conducted to verify the therapeutic effects of ZSSF on a zebrafish depression model induced by risperdal. Results 188 targets were screened to find the interactions between depression and ZSSF. The protein-protein interaction result showed that ZSSF primarily targeted TNF-α, IL-2, IL-6, IL1β, and IL-10 to produce its antidepressant effect. KEGG pathway enrichment analysis revealed that ZSSF exerted its effects on depression through various signaling pathways, including the TNF, PI3K-Akt, and cGMP-PKG signaling pathways. The result of the animal experiments showed that the treatment groups given high, medium, and low doses of ZSSF exhibited significant improvements in movement distance under acoustic and light stimulation compared with the model group(P<0. 05). The speed of movement of the treatment groups was also significantly faster(P<0. 01). Additionally,the mRNA expression levels of TNF-α, IL-2, IL-6, IL-1β, and IL-10 were up-regulated in the brain tissues of zebrafish in the high-, medium-, and low-dosage groups of ZSSF compared those in the model group ( P<0. 001). Conclusions ZSSF exerts its antidepressant effect through multiple components and targets, and its antidepressant effects may be associated with its inhibition of inflammatory factors.
