Objective To explore the mechanism of action of Neiyi Soft Extract in the treatment of endometriosis(EMS)based on network pharmacology and transcriptomics.Methods A model of SD rat with EMS was replicated by autotransplantation method.After successful modeling,the rats were randomly divided into model group,Neiyi Soft Extract low-,medium-and high-dose groups and dienogest group,with 10 rats in each group.Another sham-operated group(10 rats)was set up.After four consecutive weeks of intervention,the volume size of the endometriotic lesions was measured,and its pathological changes were detected by hematoxylin-eosin(HE)staining.RNA was extracted from the rat lesions for transcriptomic sequencing,and Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)functional enrichment analysis were performed on the differential genes among various groups.The main active ingredients and their targets of Neiyi Soft Extract were collected and screened in databases such as TCMSP,the disease targets of EMS were collected through databases such as OMIM,the intersection targets between the drug ingredient targets and the diseases were obtained by using Venn diagrams,GO enrichment analysis and KEGG pathway enrichment analysis of genes in the intersection targets of the drug ingredients and the diseases were performed by using gene enrichment analysis online tool(Metascape).The network pharmacology enrichment pathway and transcriptomics enrichment pathway were taken for intersection,the mRNA and protein expression levels of the key targets on the intersection pathway were correspondingly detected by real-time quantitative polymerase chain reaction(qPCR)and Western Blot,respectively.Results The volume of lesions in the model group was significantly increased compared with that of the sham-operated group(P<0.01);the volume of lesions in rats in the drug-administered groups was significantly reduced compared with that of the model group(P<0.05).A total of 341 active ingredients were obtained from Neiyi Soft Extract,and there were 2 178 disease-related targets of EMS,and 278 intersections between drug targets and disease targets;189 differential genes were screened in the sham-operated group and the model group;255 differential genes were screened in the model group and Neiyi Soft Extract high-dose group;and 740 differential genes were screened in the sham-operated group and the Neiyi Soft Extract high-dose group,including 390 up-regulated genes and 350 down-regulated genes.The result of intersection of KEGG enrichment pathways between the network pharmacology and transcriptomics showed that the distriution mainly included P53 signaling pathway,FOXO signaling pathway,cell cycle,etc.The qPCR and Western Blot validation results showed that Neiyi Soft Extract could inhibit the proliferation of endometrial stromal cells and up-regulated the mRNA and protein expression levels of BAX,Caspase3 in EMS rats(P<0.05 or P<0.01),and down-regulated BCL-2 mRNA and protein expression levels(P<0.05 or P<0.01).Conclusion Neiyi Soft Extract may play a therapeutic role in the treatment of EMS by regulating the P53 signaling pathway.