The purpose of this study was to characterize the putative anxiolytic-like effects of the 70%ethanol extract of Portulaca oleracea (EPO) using an elevated plus maze (EPM) in mice. The EPOwas orally administered at 50, 100, 200 or 400 mg/kg to ICR mice, 1 h before the behavioralevaluation in the EPM, respectively. Control mice were treated with an equal volume of 10%tween 80, and positive control mice with diazepam (1 mg/kg). Single treatments of the EPOsignificantly increased the percentage of time spent and arm entries into the open arms of theEPM versus controls (P < 0.05). Moreover, there were no changes in the locomotor activity andmyorelaxant effects in any group compared with the saline controls. In addition, the anxiolytic-like effects of the EPO were blocked by flumazenil (10 mg/kg, i.p), a GABAA antagonist not byWAY 100635 (0.3 mg/kg, i.p), a 5-HT1A receptor antagonist. These results indicate that P. oleraceais an effective anxiolytic agent, and suggest that the anxiolytic-like effects of P. oleracea ismediated via the GABAergic nervous system.