To explore the underlying mechanism of β-sitosterol through network pharmacology.Methods:Firstly，the 2D structure of β-sitosterol was downloaded from PubChem，and targets were predicted by PharmMapper.The results were input into UniProt to extract the gene names of targets.The disease information corresponding to the targets was retrieved from TTD.STRING was used for the construction of an association network of gene functions and DAVID for GO and pathway enrichment.Results:A total of 223 targets and 86 pieces of disease information were obtained from PharmMapper and TTD，respectively.Genes with high degrees in the association network of gene function included RAC1，B2M，ESR1，GTF2F2，AR，and KDR.Four significant items in the GO enrichment analysis were transcription initiation of RNA polymerase Ⅱ promoter，steroid receptor，steroid-mediated signaling pathway，and peptide tyrosine autophosphorylation.There were 11 significant pathways in the pathway enrichment analysis，including Rho-GTPase cycle，NRAGE signals death through JNK，neurotrophin signaling pathway，basic transcription factor，MyD88 deficiency(TLR2/4)，IRAK4 deficiency(TLR2/4)，DAP12 signaling pathway，β-alanine metabolism，oocyte meiosis，Rap1 signaling pathway，and MyD88:MAL(TIRAP) cascade initiated on the plasma membrane.Conclusion:The findings of this study reveal that β-sitosterol can act on various diseases including nervous system，reproductive system，immune system，infectious disease，and tumors through multi-target and multi-pathway.