The present study aimed to investigate the role of hydrogen sulphide (H₂S) in the cardioprotection induced by ischemic postconditioning and to examine the underlying mechanisms. Cardiodynamics and myocardial infarction were measured in isolated rat hearts. Postconditioning with 6 episodes of 10 sec ischemia (IPostC) significantly improved cardiodynamic function, which was attenuated by blockade of endogenous H₂S production with DL-propargylglycine. Moreover, IPostC significantly stimulated H₂S-synthesis enzyme activity during the early period of reperfusion. However, DL-propargylglycine only attenuated IPostC-induced activation of PKC and PKC but not that of PKC, Akt and eNOS. These data suggest that endogenous H₂S contributes partially to the cardioprotection of IPostC via stimulating PKC and . Postconditioning with 6 episodes of 10 sec infusion of NaHS (SPostC) or 2 min continuous NaHS infusion (SPostC2) stimulated activities of Akt and PKC, improved the cardiodynamic performances and reduced myocardial infarct size. Blockade of Akt with LY294002 (15 µM) or PKC with chelerythrine (10 µM) abolished the cardioprotection induced by H₂S postconditioning. SPostC2, but not SPostC, also additionally stimulated eNOS. We conclude that endogenous H₂S contributes to IPostC-induced cardioprotection. H₂S postconditioning confers protective effects against ischemia-reperfusion injury through activation of Akt, PKC and eNOS pathways.