Objective To investigate effects of Mosla chinensis seed oil on physiological behavior and antioxidant index in Drosophila melanogaster. Methods One-day-old wild-type Drosophila melanogaster was divided into control group, treatment group and model group. Drosophila model group was deprived of sleep by repetitive light stimulation at night to establish insomnia model, while the control group was exposed to the basal culture data. The negative geotaxis ability was used to explore the optimal concentration and period of drug treatment, and appetite test, negative geotaxis ability, stress test, olfactory memory test and detection of sleep-wake rhythm were used to explore the effects of Mosla chinensis seed oil on physiological behavior. In addition, activities of Super Oxidase Dimutase (SOD), Catalase (CAT) and Propanedial (MDA) were detected by enzyme-linked immunosorbent assay (Elisa). Results The Mosla chinensis seed oil enhanced the locomotion ability of 30-day-old Drosophila melanogaster (P < 0.01). Increased the activity of SOD and CAT (P < 0.01%) and decreased the content of MDA(P < 0.01); The sleep time of Drosophila was prolonged (P < 0.05 or P < 0.01%);Improved olfactory memory in fruit flies.After sleep deprivation, the night sleep time of female Drosophila model group decreased (P < 0.05%), and that of male drosophila model group decreased (P < 0.01). After feeding Mosla chinensis seed oil, the night sleep time of sleep deprived female drosophila was extended (P < 0.05), and that of male drosophila was extended (P < 0.01%). Conclusions Using Drosophila as model animal, it was found that Mosla chinensis seed oil had certain antioxidant ability in vivo, prolonging sleep time and improving olfactory memory of drosophila sleep-deprived drosophila.