Oleuropein is one of the main phenolic secoiridoid of the olive leaf extract, which is known for its antioxidant and anti-inflammatory effects. The main objective of the present study was to investigate the effectiveness of oleuropein in the ulcerative colitis treatment. An experimental study was designed on rats, which were divided into three groups, group 1 (normal control), group 2 (induced for ulcerative colitis and untreated), and group 3 (induced for ulcerative colitis and treated with oleuropein). Colonic tissue samples were collected from all studied groups and the oxidative stress and antioxidant activity were assessed by evaluating malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), myeloperoxidase (MPO), and nitric oxide (NO) levels. The expression levels of pro-inflammatory cytokines such as IL-1β, TNF-α, IL-10, COX-2, iNOS, TGF-β1, MCP-1, and NF-κB, the pro-apoptotic gene Bax, and the anti-apoptotic gene Bcl2 were assessed in colon tissues to evaluate the effectiveness of oleuropein treatment. Oleuropein was effective on reducing the mortality rate and disease activity index. Oleuropein caused a significant reduction in colon MDA, MPO, and NO levels and a significant elevation in SOD, CAT, and GPX levels and induced the down regulation of analyzed proinflammatory cytokines. Also, downregulation of Bax and upregulation of Bcl2 were observed as a result of oleuropein treatment in comparison with untreated acetic acid induced ulcerative colitis group. Oleuropein showed intestinal anti-inflammatory, antioxidant, and anti-apoptotic effects in ulcerative colitis experimental model.