Aim To search for global protein changes in ischemic preconditioning (IP) using proteomic technology. Methods The SD rats were divided into two groups including IP group and sham groups. The left ventricle tissues of preconditioned and sham groups were directly sampled for proteomic analysis. Results Analysis of two dimensional electrophoresis (2-DE) showed: 704±27 protein spots were seen in Sham ventricle gels and 778±35 protein spots in IP ventricle gels. Differential analysis using the Student’s t-test (p<0.05) showed that the expression of 23 protein spots changed, 12 protein spots were identified by mass spectrometry. Conclusions Myocardial IP resulted in the changes of protein expression profiles in the myocardium. The differential proteins might function as molecular chaperone, decrease free radical and promote the energy metabolism of myocardium to confer cardioprotection.