Zearalenone (ZEA) induces oxidative damage in porcine endometrial stromal cells (ESCs), which is a critical factor affecting the growth and reproduction of female pigs. We hypothesize that rutin, a flavonoid antioxidant, can alleviate ZEA-induced cellular damage through the p53 signaling pathway. In this experiment, porcine ESCs were used as a research model. After transfection with siRNA to silence the p53 gene, rutin and ZEA were added to the cocultured porcine ESCs. Rutin notably reduced the ZEA-induced apoptosis rate, ROS, MDA, and 4-HNE levels via the p53 pathway; lowered the expression of LC3, Beclin-1, p62, Bax, and CHOP; and significantly improved cell viability, GSH-Px, T-SOD enzyme activity, and Bcl-2 expression. It also inhibited both apoptosis and autophagy. These findings suggested a novel antioxidant mechanism by which rutin mitigated the toxic effects of ZEA, highlighting the role of the p53 gene knockdown in this process.