Objective To investigate the effects of citicoline combined with rehabilitative training on motor function after focal cerebral ischemia. Methods One hundred and twenty male adult Sprague-Dawley rats ( 3 months old) were subjected to left middle cerebral artery occlusion (MCAO) by suturing. Ninety-six of them were randomly divided into four groups of 24: a control group, a drug group, a rehabilitative training group, and a drug combined with rehabilitative training group. The rats in the control group did not receive any treatment. Three days after reperfusion, those in the drug group received 500 mg/kg of citicoline daily; those in the rehabilitative training group received motor training including balancing, grasping, rotating and walking exercises; those in the combined group received both citicoline and the motor training program. Behavioral tests were administered at the 7th, 14th and 21st days after MCAO. At the same time points, immunohistochemistry was used to detect calcium-binding protein S100 and β-tubulin expression in the peri-ischemia region of the cortex. Results ①As compared with the control group, the drug group at the 7th, 14th and 21st day after MCAO and the other two groups at the 7th day after MCAO showed no significant difference in average behavior scores. But at the 14th and 21st day the rehabilitative training group as well as the drug combined with rehabilitative training group had average behavior scores significantly superior to those of the control group, especially in the drug combined with rehabilitative training group. ②As compared with the control group, the expression of S100 and β-tubulin in the drug and rehabilitative training groups at the 7th day after MCAO showed no significant difference. In the drug combined with rehabilitative training group the average score was significantly higher than in the other groups at the 7th day after MCAO. All the other groups had average scores significantly higher than that of the controls at the 14th and 21st day after MCAO, especially the drug combined with rehabilitative training group. Conclusions Citicoline combined with rehabilitative training can significantly improve motor function recovery in rats. The effect of the combined modalities was more obvious, and the functional enhancement might be partially attributable to the up-regulation of S100 and β-tubulin in the cortex.