Objective To explore whether the cAMP-PKA-pCREB signal pathway plays a role in promoting the recovery of motor function after rehabilitation training in cerebral ischemia-reperfusion rats. Methods The middle cerebral artery occlusion model (MCAO) was established by modified Longa nylon occlusion method in adult male Sprague-Dawley rats. The 84 MCAO rats were selected and randomly assigned to four groups: the natural recovery group without any special training (group B, n=24),natural recovery group with Rp-cAMP (group C, n=24), rehabilitation training group (group D, n=18) and rehabilitation training with Rp-cAMP (group E, n=18), and in addition a control group (group A, n = 12). To establish rat MCAO models immediately after injection of Rp-cAMP into the lateral ventricle of the brain. The rats in the groups D and E were trained by balance beam, bar rotating and rolling exercises started at 48 h after MCAO. The expression of PKA was determined by enzyme-linked immunosorbent assay (ELISA) and the pCREB protein expression was detected by Western blot assay. Motor function was assessed by balance beam test. Results (1) The motor function score in the group C was significantly higher than that of group B, suggesting that Rp-cAMP inhibited the recovery of motor function in the cerebral ischemia-reperfusion rats. The score of group D was significantly lower than that of groups B and E, indicating that Rp-cAMP inhibited the promoting effect of rehabilitation training on motor function in the cerebral ischemia-reperfusion rats. (2) The expressions of PKA and pCREB proteins detected at 2nd, 7th, 14th, and 21th days after surgery showed that their expressions in the group D were significantly higher than those of the groups B and E, indicating that rehabilitation training promoted the expression of PKA and pCREB, and Rp-cAMP significantly inhibited the promoting effect of rehabilitation training on the expressions of PKA and pCREB proteins. Conclusion cAMP/PKA-pCREB signal transduction pathway may mediate a promoting effect of rehabilitation training on the recovery of motor function after ischemic stroke in rats.