Introduction: Neuroprotective effect of melatonin in traumatic brain injury (TBI) has been shown in many animal studies. This clinical trial was designed to investigate the effect of melatonin on neurological outcome in diffuse axonal injury (DAI). Methods and Materials: Seventy male patients 18-60 years with moderate to severe DAI admitted within the first 4 hours of injury were randomly divided into control and melatonin groups. The melatonin group received orally 3 mg melatonin at the time of admission, and itcontinued until the 15th day consecutive in addition to standard treatment. Glasgow coma scale (GCS) assessment was performed at the time of admission, fifteen days after injury and the day of discharge, while functional independence measure (FIM) and Glasgow coma scale (GOS) at the time of admission and the third and sixth months after injury. Results: There was significant difference in GCS between the control and melatonin groups at fifteen days post injury and GCS increased in the melatonin group (P<0.01). There was an increase in GOS and FIM scores in the melatonin group compared to the control group at three and six months after DAI (P<0.05). This score increased in the melatonin group at six months after injury compared to three months after injury (P<0.001). Unlike the control group, no death occurred in the melatonin group. Conclusion: The findings of this study showed the treatment of melatonin could probably improve neurological outcome in male patients with DAI. A study with larger sample size is needed to confirm the efficacy of melatonin in DAI. The mechanism of the effect of melatonin in DAI will be examined in a future study.