Hydroxytyrosol (HYT) is one of the main alcoholic compounds of the olive leaves extract (OLE), which is known for its beneficial effects. This study aimed to investigate the effectiveness of olive leaves extract standardized with 25% hydroxytyrosol (OLES-25%HYT) in treatment of induced ulcerative colitis. Three groups of albino rats, were divided as following, group 1 (normal control), group 2 (induced ulcerative colitis and untreated) and group 3 (induced ulcerative colitis and treated with OLES-25%HYT). Colonic tissue samples were collected from all studied groups, the antioxidant activity for malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), myeloperoxidase (MPO) and nitric oxide (NO) were performed. The expression of pro-inflammatory cytokines, the apoptotic gene Bax and the anti-apoptotic gene Bcl2 was obtained in colon tissue to evaluate the OLES-25%HYT effect on ulcerative colitis. OLES-25%HYT showed effectiveness on reduction of mortality rate and disease activity index (DAI). Also, reduced oxidative stress and inflammation in colon tissue, OLES-25%HYT showed a significant reduction in colon MDA, MPO and NO levels and a significant elevation in SOD, CAT and GPX levels and cause down regulation of pro-inflammatory cytokines. Also, the apoptotic gene Bax downregulated and the anti-apoptotic gene Bcl2 upregulated as a result of the treatment compared to untreated induced ulcerative colitis group. OLES-25%HYT showed intestinal anti-inflammatory, antioxidants and anti-apoptotic effects in experimental models of ulcerative colitis.