Abstract: Objective To investigate the effect of resveratrol on endometrial cancer based on network pharmacology, molecular docking and experimental verification.Methods The target of resveratrol was retrieved from HERB and ECTM databases, and the target of endometrial cancer was obtained from Genecards database. The intersection genes of resveratrol for the treatment of endometrial cancer were imported to the STRING database to establish a drug-disease-target network by Cytoscape software. Then, GO and KEGG pathway enrichment analyses were performed for intersection targets using OmicShare cloud platform. Resveratrol and hub targets were analyzed by molecular docking. Furthermore, a 1-methyl-3-nitro-1-nitrosoguanidine(MNNG)-induced EC model of mice was constructed and the mice were randomly divided into five groups: a Control group, a Res group, a MNNG group, a MNNG + Res group, and a MNNG + Res + MAPK/ERKi group. The protein levels of ERK and p-ERK in mouse uterus were detected by Western blot. The contents of 15 estrogen active substances in serum and endometrial tissue of mice were detected by LC-MS/MS.Results A total of 174 intersection targets were obtained for the treatment of endometrial cancer by resveratrol. KEGG enrichment signaling pathways included PI3K-Akt signaling pathway and MAPK signaling pathway. The top ten hug targets were screened and resveratrol showed stable binding activity with MAPK3, according to molecular docking. Moreover, compared with the Control group, the Res group, MNNG+Res group and MNNG+Res+MAPK/ERKi group showed increased phosphorylated ERK levels in endometrial tissues. According to LC-MS/MS analysis, the MNNG group showed significantly decreases in the contents of 2-MeOE1, 2-MeOE2 and 4-MeOE1 in serum and uterine tissue and increases in the content of 4-OHE2(P<0.05). Compared with the MNNG group, the MNNG+Res group presented remarkable increases in the levels of 4-MeOE1 in serum as well as 2-MeOE1 and 2-MeOE2 in endometrial tissue, but decrease in the content of 4-OHE2 in serum and endometrial tissues(P<0.05). In the MNNG+Res +MAPK/ERKi group, the effect of resveratrol on the reversal of estrogen homeostasis imbalance was obviously weakened.Conclusions Resveratrol is characterized by multiple targets and multiple approaches for the treatment of endometrial cancer. It regulates estrogen metabolism by activating the MAPK pathway, reduces the accumulation of hydroxylated estrogen in endometrial tissue, and restores endometrial estrogen homeostasis, in order to prevent the occurrence of endometrial cancer.