Ischemic postconditioning (IP) has been shown to provide protection for ischemia/reperfusion (IR) injury, but its efficacy is limited. In this study we hypothesized that spontaneous running wheel (RW) could improve neuroprotection efficacy of IP for IR. We established mouse models of IR and showed that compared to Sham group, IR group had obvious brain infract and neurological dysfunction. In IR+IP group, brain infract and neurological dysfunction improved compared to IR group. However, in IR+IP+RW group brain infract and neurological dysfunction improved much better. TUNEL assay showed that IP but not RW significantly reduced the number of apoptotic cells after IR. However, the number of apoptotic cells was significantly reduced in RW+IP group. In addition, the levels of pro-apoptotic factors increased in IR group but significantly reduced in IR+IP+RW group, while the levels of anti-apoptotic factors decreased in IR group but significantly increased in IR+IP+RW group. Moreover, in IR+IP+RW group, MDA level was further decreased and SOD level was further increased compared to IR+IP group. Finally, both PI3K inhibitor and STAT3 inhibitor significantly worsened brain infract and neurological dysfunction and promoted apoptosis in IR mice. In conclusion, RW combined with IP reduces brain infract and neurological dysfunction in mice after IR, and this is associated with enhanced anti-apoptotic and anti-oxidant benefits via the activation of PI3K and STAT3 pathways.