OBJECTIVES Jujubae fructus (JF, <i>Ziziphus jujuba</i> var. inermis Rehder) is an economically and agriculturally significant perennial fruit tree crop of the Rhamnaceae family and is used as a traditional herbal medicine. However, the anti-fatigue effect of JF has not been reported yet. The purpose of this study was to examine the anti-fatigue effect of JF by in vivo experiments.Methods Male ICR mice were classified into a saline-treated swimming load test group (control) and JF (1, 10, 100 mg/kg) supplied swimming load test group (JF), and randomly assigned to 5 mice per group. Distilled water and JF were orally administered. The anti-fatigue effect of JF was evaluated by open field test (OFT) and forced swimming test (FST).Results JF significantly increased the total distance moved in OFT in a dose-dependent manner. JF was significantly decreased immobility time compared to control group in the FST. The increase in lactic acid and lactate dehydrogenase after FST was suppressed by JF treatment. Consumption and recovery of energy sources (free fatty acid and triglyceride) were improved after FST through JF treatment. Administration of JF decreased the expression of neurotransmitters such as norepinephrine (NE), serotonin (5-HT), 5-hydroxyindole-acetic acid (5-HIAA), dopamine (DA) by FST in brain stem and hypothalamus.Conclusions Based on these results, this study provides scientific proof for anti-fatigue effect of JF and it could be a useful material.