Propolis has a broad spectrum of biological activities; however, whether its essential oils have neuroprotective effects is unknown. In this study, we found that propolis essential oil (PEO) could significantly reverse the anxiety-like behavior of restraint-stressed mice, and has no effect on locomotor activity. Furthermore, PEO significantly decreased the plasma levels of cortisol (CORT), adrenocorticotropic hormone (ACTH) and malondialdehyde (MDA), whereas it increased the activity of superoxide dismutase (SOD) in restraint-stressed mice. These results strongly suggest that PEO has therapeutic effects on anxiety through antagonizing the hyperfunction of hypothalamic-pituitary-adrenal (HPA) axis and improving the ability of antioxidation in brain tissue.