To explore the mechanism of immunity enhancement by Polygonati Rhizoma.Methods:The active components of Polygonati Rhizoma were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)，and SwissTargetPrediction was used to predict the corresponding target genes.GeneCards and DrugBank were employed to screen out the target genes of immunosuppression(IS).The Venn diagram was established to obtain the common genes shared by Polygonati Rhizoma and IS.String was employed to construct the protein-protein interaction network，and Cytoscape was used to construct the “Polygonati Rhizoma-active component-target gene-IS” network.Metascape was used to conduct gene ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses on the common genes.Molecular docking was adopted to verify the binding between active components and targets.The RAW264.7 cell model induced by lipopolysaccharide(LPS) was established.The methyl thiazolyl tetrazolium(MTT) method was used to examine the toxicity of Polygonati Rhizoma extracts at different concentrations to cells.The Griess method was used to test the effect of Polygonati Rhizoma on nitric oxide(NO) in cells.Results:ESR1，SRC，IGF1R，PTGS1，and PTGS2 were predicted to be involved in key biological processes and pathways.GO enrichment showed that cell activation，inflammatory response，and cell response to stress were the main biological processes involved.KEGG enrichment showed that the AGE-RAGE signaling pathway in diabetic complications，AMPK，and nuclear factor-kappa B(NF-κB) signaling pathways were the critical pathways.Molecular docking results showed strong binding affinity between active components and core targets.Compared with the LPS group，the Polygonati Rhizoma extract groups(50，200，and 800 μg/mL) showed reduced production of NO in the RAW264.7 cells(P<0.01).Conclusion:This study preliminarily predicted the targets such as ESR1 and SRC and signaling pathways such as AMPK and NF-κB of Polygonati Rhizoma in enhancing immunity.Polygonati Rhizoma can enhance immunity by regulating the release of NO in RAW264.7 cells.