Background: Ulcerative colitis (UC) is a chronic inflammatory disease with increasing incidence and prevalence worldwide. Currently used treatments of UC are unsatisfactory while natural biactive compounds are considered to be emerging therapeutic agents. Luteolin (Lut) is a natural compound with beneficial effects in a variety of diseases, however, its effect in UC has been poorly studied. Purpose: This study attempted to clarify molecular mechanisms behind the biological activity of Lut in the treatment of UC, with the emphasis on the extracellular signal-regulated kinase (ERK) signaling pathway. Study design: The effect of Lut was investigated in a posttreatment and cotreatment of dextran sulfate sodium (DSS)- induced experimental colitis in mice. In addition, the role of ERK1/2 in the mechanism of action of Lut in experimental colitis was investigated using ERK1/2 inhibitor. Methods: Clinical symptoms (disease activity index) and histopathological changes of the colon were evaluated during disease development. The expression of mitogen-activated protein kinase (MAPKs), inflammatory markers and apoptotic and autophagic proteins were determined by western blot. Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 expression and cellular localization were analyzed by immunohistochemistry. Results: Lut administration attenuated symptoms of DSS- induced colitis in mice, ameliorated colon tissue damage and reduced inflammation, apoptosis and autophagy. The effect was more pronounced if Lut was administrated at the time of colitis induction (cotreatment). The administration of ERK1/2 inhibitor exacerbated DSS-induced colitis symptoms and prevented the protective effects of Lut. Conclusion: This study provides new mechanistic details underlying the anti-inflammatory, anti- apoptotic and anti-autophagic effects of Lut through the activation of the ERK signaling pathway. This suggested that Lut can be used as a novel therapeutic candidate in the treatment of UC or could be used as a supplement to existing therapy.