Background: The ability of pharmacological preconditioning mimetics to confer long lasting and sustained cardioprotection may be logical criterion to develop a drug which can be used clinically for cardioprotection. We propose here that the use of long acting PDE5A inhibitor, tadalafil may confer sustained cardioprotection against ischemia. Methods: Tadalafil (5 mg/kg) was administered orally to male C57 Black/6J mice (n=6 in each treatment sub-group at each time point studied). Hearts were isolated and subjected to 40-min ischemia and 30-min reperfusion on Langendorff's apparatus at 1-h, 12-h, 24-h, 36-h, 48-h, 60-h, 72-h and 108-h after tadalafil administration. In 1-h to 48-h sub-groups, tadalafil was given once at 0-h only. In 60-h and 72-h sub-groups, tadalafil was given twice at 0-h and 36-h. Similarly, in 108-h sub-group, tadalafil was administered at 0-h, 36-h and 72-h. In the same sub-groups, Wortmannin (Wort, 15 µg/kg IP), an inhibitor of PI3K, or 5-hydroxydecanoic acid (5-HD, 5mg/kg IP), an inhibitor of mitoKATP channels, was given together with tadalafil and the hearts were subjected to ischemia/perfusion at 36-h, to determine whether tadalafil effect on ischemia/reperfusion injury was abolished. Results: Tadalafil treatment reduced left ventricular end diastolic pressure and increased left ventricular developed pressure as well as reduced LDH release. This protection remained till 36-40 hrs and thereafter it vanished. Readministeration of tadalafil at 36 hrs and 72 hrs restored the protection till 108 hrs. Tadalafil treatment accelerated Akt phosphorylation in cardiac tissue and decreased myocyte apoptosis. Administration of wort abolished the beneficial effects of tadalafil on hemodynamic parameters and myocyte apoptosis, together with significantly reduced Akt phosphorylation. 5-HD also abolished the anti-apoptotic effect of tadalafil. Conclusion: It is concluded that tadalafil treatment induces long term protection of ischemic myocardium via PI3K/Akt signaling pathway