Background & Objective: In Previous studies have shown that antioxidant properties of spinach play a role in age-related declines in neuronal and cognitive function and neurodegenerative diseases. This study was designed to evaluate anxiolytic effects in different doses of the aqueous extracts of Spinach leaves.Materials & Methods: In this study, fifty male mice (25-30 g) in model EPM were used. Different doses of the extract (0.05, 0.10 and 0.15 g/mice) were given to 3 test groups for 1 month. Also, water and sucrose 3% (w/v) were given to 2 control group for 1 month. At the first time for increasing activity, animals have put inside the black wall box for 5 min. Then animal transfers to the elevated plus maze and evaluated standard indexes of anxiety reaction.Results: Results showed that % open time and the number of open arm entries was significantly longer in doses (0.10 and 0.15), (0.15) g of spinach extract than control group, respectively (P<0.05).Furthermore, no significant differences were found in the number of total arm entries between groups. In comparison with control group, the number of SAP was significantly shorter in doses 0.10 and 0.15 g of spinach extract (P<0.05).Conclusion: It is concluded that aqueous extract of spinach could be decrease anxiety in higher doses.