Intestinal oxidation-reduction balance is essential to protect host health. Research on the underlying mechanisms of oxidation homeostasis can provide conditions to the treatment or protection of intestinal oxidative stress. Therefore, the mechanism of polysaccharide (MBP) with alkali-extracted from mung bean skin in alleviating oxidative damage induced by H2O2-induced in intestinal epithelial cells (IEC)-6 cells was explored. Results indicated that MBP effectively mitigated the H2O2-induced decrease in IEC-6 cell viability, restored superoxide dismutase (SOD) activity, lowered malondialdehyde (MDA) content, and decreased reactive oxygen species (ROS) level. Western blotting showed that MBP may mitigate oxidative damage through NF-κB, MAPK and Nrf2 signaling pathway, and RNA sequencing (RNA-seq) analysis confirmed that MBP can protect cells by regulating MAPK and NF-κB signaling pathways. Moreover, the results of the inhibitor experiment also support the above hypothesis. These results verified that MBP has the capacity to mitigate oxidative damage induced by H2O2 in IEC-6 cells through MAPK and NF-κB signaling pathway. The study showed that MBP can assist preventing intestinal diseases relating to oxidative stress, which could facilitate the development of functional foods.