Dihydromyricetin (DHM), the major bioactive flavonoid ingredient extracted from the leaves of Ampelopsis grossedentata (Hand.-Mazz) W.T. Wang displays multiple pharmacological activities, including oxidation resistance, anti-tumour properties and free radical scavenging capacities. However, the role of DHM in sleep deprivation (SD)-induced memory impairments and its underlying molecular mechanisms are unclear. The aim of the present study was to evaluate the effects of DHM on oxidative stress and its role in ameliorating memory impairment induced by acute SD. DHM (100, 50, 25 mg/kg) and melatonin (10 mg/kg) were administered to mice via oral gavage. The open field test was used to evaluate motor function. Spatial learning and memory were assessed using the Morris water maze task. Malondialdehyde, glutathione, and glutathione disulfide levels, as well as superoxide dismutase enzyme activity, were assessed to determine the level of oxidative stress. In addition, we employed quantitative real-time PCR assays to examine the gene expression of 29 key proteins, including protein kinase A (PKA), cAMP response element binding protein (CREB), and adcy1. The levels of proteins including those of GABA B R S , GABA A Rα5, GluR1, BDNF and PSD95, were detected by western blotting. The results showed that DHM significantly attenuated SD-induced spatial learning and memory impairments (P < 0.01). The possible underlying mechanisms of DHM may be attributed to its ability to reduce oxidative stress and restore synaptic plasticity.