Survivin is one of the inhibitors of the apoptosis gene family that has been implicated in both inhibition of apoptosis and mitosis regulation. In this study, using a protein delivery system, we demonstrated the potential cytoprotective effect of survivin as a therapeutic protein in rat embryonic cardiac myoblasts (H9c2 cells) and acute doxorubicin(DXR)-induced cardiomyopathy rat model (a single intraperitoneal injection of DXR, 15 mg/kg). We constructed a recombinant survivin fused to the protein transduction domain (PTD) derived from HIV-1 TAT protein. Purified recombinant TAT-survivin protein was efficiently delivered to H9c2 cells, and its transduction showed cytoprotective effect against doxorubicin. Moreover, transduction of TAT-survivin also attenuated DXR-induced apoptosis, which was accompanied by reduced caspase-3 activity. TAT-survivin also blocked the p53 activation, Bax translocation, cytochrome C release and activation of caspase cascade induced by doxorubicin in H9c2 cells. However, the survivin mutant protein at phosphorylation site failed to protect doxorubicin-induced H9c2 cell death. In addition, intraperitoneal injection of TAT-survivin into rat resulted in efficient protein transduction in heart tissues and preserved heart function in Langendorff perfusion model of DXR-induced cardiomyopathy rat, as determined by 20 minutes of global no-flow ischemia, followed by 40 minutes of reperfusion. These results suggest that the PTD-mediated delivery of survivin pretreatment may offer significant protection against DXR-induced myocardial damage.