Although the effect of mood stabilizer valproic acid (VPA) through multiple signaling pathways has been shown, its therapeutic mechanism is still largely unknown. We investigated the effect of VPA (200 mg/kg, every 12 h) in sleep deprivation (SD) rats (72 h), the manic-like animal model, focusing on the N-methyl-D: -aspartic acid (NMDA) receptor and signaling mediators of synaptic plasticity such as extracellular signal-regulated protein kinase (ERK), cAMP response element-binding protein (CREB), B cell chronic lymphocytic leukemia/lymphoma 2 (BCL2), and brain-derived neurotrophic factor (BDNF). SD reduced the expression of the NR2B subunit of the NMDA receptor in the frontal cortex and hippocampus but did not affect the expression of NR1 and NR2A subunits. In comparison, VPA inhibited the SD-induced reduction of NR2B expression in both brain regions. In addition, SD attenuated ERK phosphorylation in the frontal cortex and hippocampus, whereas VPA prevented the attenuation. VPA also protected the SD-induced decrease of CREB phosphorylation, BCL2 expression, and BDNF expression in the frontal cortex but not in the hippocampus. These results indicate that VPA could regulate NMDA receptor-ERK signaling in SD rats, preventing the SD-induced decrease of the expression of NR2B subunit and the activation of ERK signaling mediators such as ERK, CREB, BCL2, and BDNF.