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In the weaning period, piglets often face oxidative stress, which will cause increased diarrhea and mortality. Genistein, a flavonoid, which is extracted from leguminous plants, possesses anti-inflammatory and antioxidative bioactivities. However, little is known about whether genistein could attenuate the oxidative stress that occurs in porcine intestinal epithelial cells (IPEC-J2). Herein, this experiment was carried out to investigate the protective effects of genistein in the IPEC-J2 cells oxidative stress model. Our results disclosed that H₂O₂ stimulation brought about a significant diminution in catalase (CAT) activity and cell viability, as well as an increase in the levels of reactive oxygen species (ROS) in IPEC-J2 cells (<i>p</i> < 0.05), whereas pretreating cells with genistein before H₂O₂ exposure helped to alleviate the reduction in CAT activity and cell viability (<i>p</i> < 0.05) and the raise in the levels of ROS (<i>p</i> = 0.061) caused by H₂O₂. Furthermore, H₂O₂ stimulation of IPEC-J2 cells remarkably suppressed gene level <i>Nrf2</i> and <i>CAT</i> expression, in addition to protein level Nrf2 expression, but pretreating cells with genistein reversed this change (<i>p</i> < 0.05). Moreover, genistein pretreatment prevented the downregulation of occludin expression at the gene and protein level, and <i>ZO-1</i> expression at gene level (<i>p</i> < 0.05). In summary, our findings indicate that genistein possesses an antioxidant capacity in IPEC-J2 cells which is effective against oxidative stress; the potential mechanism may involve the Nrf2 signaling pathway. Our findings could offer a novel nutritional intervention strategy to enhance the intestinal health of piglets during the weaning process.