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A single dose of MTX (20\u2009mg\u2009kg<jats:sup>\u22121<\/jats:sup>, i.p.) was followed by i.p. saline or melatonin injections (10\u2009mg\u2009kg<jats:sup>\u22121<\/jats:sup>, MTX\u2009+\u2009Mel) for the next 5 days. On the fifth day, intestinal transit was assessed using charcoal propagation. Rats were decapitated and small intestinal segments were fixed for light (LM) and scanning electron microscope (SEM) examinations. Other intestinal segments were stored to measure glutathione (GSH) and malondialdehyde (MDA) levels, myeloperoxidase (MPO) and ATPase activity. MTX led to loss of more than 10% of the initial body weight (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.01). Conversely, weight loss was markedly less in the melatonin\u2010treated MTX group (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05). Bowel motility was increased in MTX\u2010treated rats, while the transit index in the MTX\u2010Mel group was not different from the control group. MTX caused decreases in GSH levels and ATPase activity, with increases in MDA levels and MPO activity. These changes were reversed in MTX\u2010Mel\u2010treated rats (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05\u2013<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.001). LM and SEM in the MTX group revealed desquamation of surface epithelium and glandular degeneration, while the epithelium was slightly damaged in the MTX\u2010Mel group. In conclusion, the present study demonstrates that melatonin is capable of reversing MTX\u2010induced intestinal dysfunctions, indicating that it may be beneficial in ameliorating the symptoms of chemotherapy\u2010induced enteritis. 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