To explore and compare the mechanism of Yupingfeng Powder and melatonin in regulating the immune system,and verify the core targets with the aging model of mice.Methods:The targets of Yupingfeng Powder and melatonin were searched in Symptom Mapping(SymMap) and Comparative Toxicogenomics Database(CTD),respectively.The ClueGO plug-in of Cytoscape was used for the gene ontology(GO) annotation of immune system process(ISP).The related targets were extracted,and the key targets were screened out.STRING was employed to build the protein-protein interaction(PPI) network of the key targets.The ClueGO plug-in was used for KEGG pathway enrichment analysis of the key targets.Eighty-four male SPF-grade mice aged 8 weeks were administrated with D-galactose at 1 000 mg/(kg·d) for 4 weeks.After that,they were assigned into three temperature groups:normal temperature group,temperature dropping group,and temperature rising group.Each temperature group was further assigned into a blank control group,a Yupingfeng Powder group,a melatonin group,and a Yupingfeng Powder+melatonin group.After the intervention,blood was collected from the eyeballs for the determination of serum levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-alpha(TNF-α) by enzyme-linked immunosorbent assay(ELISA).Results:A total of 339 targets and 325 targets were obtained for Yupingfeng Powder and melatonin,respectively.GO annotation predicted 11 and 32 ISPs for Yupingfeng Powder and melatonin,which involved 62 and 84 targets,including 20 and 32 key targets,respectively.The P-values of PPI networks constructed with key targets were less than 3.33E-16 and 1.0E-16,respectively,with significant statistical significance.KEGG pathway enrichment showed that the key targets of Yupingfeng Powder and melatonin were mainly involved in 33 and 61 signaling pathways,respectively.In the experiments verifying the effects of Yupingfeng powder and melatonin on serum IL-1β,IL-6,and TNF-α levels in the elderly mice under different air temperatures,the positive results all ranked top in the topological analysis of targets.Conclusion:Both Yupingfeng Powder and melatonin can regulate the immune system through complex pathways and targets.They target similar pathways targeted but different targets.The combination of the two has certain advantages in clinical practice.