Introduction: Stroke is a major cause of mortality and long-term disability among adults. The risk of stroke is rapidly increasing in women after menopause. It has been reported that exercise reduces ischemia and reperfusion injury in rat model of stroke. The aim of this study was to investigate the effect of exercise on stroke outcome in the permanent middle cerebral artery occlusion in ovariectomized mice. Materials and Methods: A group of 32 female mice (25-35g) were randomly divided into 4 groups as following (8 mice in each group): ovariectomy+stroke, stroke, ovariectomy+exercise+stroke, and sham. Seven days before exercise preconditioning, mice were ovariectomized. The exercise group was forced to run on a treadmill 5 days per week, for 40 min/day at a speed of 18 m/min for four weeks. Stroke was induced by permanent middle cerebral artery occlusion (MCAO) method five weeks after ovariectomy. The infarct volume, sensory-motor deficits, and neurological deficits were studied. Results: Infarct volume in the ovariectomy+exercise+stroke and stroke groups was significantly smaller compared to ovariectomy+stroke group. In ovariectomy+exercise+stroke and stroke groups, neurological deficits were significantly lower than ovariectomy+stroke gorup, respectively. sensory-motor deficits were also lower in the ovariectomy+exercise+stroke and stroke groups compared to ovariectomy+stroke group. Conclusion: The present data suggest that exercise preconditioning plays a neuroprotective role in ovariectomized animals and improves stroke outcome in a permanent model of MCAO.