This study investigates the role of the xenobiotic metabolism factor, pregnane X receptor (PXR), in mediating deoxynivalenol (DON)-induced hepatocellular oxidative stress. The PXR agonist 3-indole-propionic acid (IPA) is found to alleviate DON-induced oxidative stress and liver injury both in vitro and in vivo. Mechanistically, IPA directly transactivates the m6A demethylase FTO, leading to site-specific demethylation and decreased abundance of YTHDC1-bound Malat1 lncRNA at single-nucleotide resolution. This reduction in m6A modification of Malat1 lncRNA reduces its stability and enhances antioxidant pathways governed by NRF2, thereby mitigating DON-induced liver injury. Malat1 knockout mice exhibit decreased DON-induced liver injury, emphasizing the role of Malat1 lncRNA in oxidative stress. Collectively, the findings establish that PXR-mediated m6A-dependent Malat1 lncRNA expression determines hepatocyte oxidative stress via m6A demethylase FTO, providing valuable insights into the potential mechanisms underlying DON-induced liver injury and offering potential therapeutic strategies for its treatment.This study investigates the role of the xenobiotic metabolism factor, pregnane X receptor (PXR), in mediating deoxynivalenol (DON)-induced hepatocellular oxidative stress. The PXR agonist 3-indole-propionic acid (IPA) is found to alleviate DON-induced oxidative stress and liver injury both in vitro and in vivo. Mechanistically, IPA directly transactivates the m6A demethylase FTO, leading to site-specific demethylation and decreased abundance of YTHDC1-bound Malat1 lncRNA at single-nucleotide resolution. This reduction in m6A modification of Malat1 lncRNA reduces its stability and enhances antioxidant pathways governed by NRF2, thereby mitigating DON-induced liver injury. Malat1 knockout mice exhibit decreased DON-induced liver injury, emphasizing the role of Malat1 lncRNA in oxidative stress. Collectively, the findings establish that PXR-mediated m6A-dependent Malat1 lncRNA expression determines hepatocyte oxidative stress via m6A demethylase FTO, providing valuable insights into the potential mechanisms underlying DON-induced liver injury and offering potential therapeutic strategies for its treatment.