Abstract: Objective To explore the role of Akt/Gsk-3β signaling pathway in sulforaphane (SFN) protecting rats from cold myocardial ischemia-reperfusion injury during heart transplantation. Methods Sixty-four health male Sprague-Dawley (SD) rats were randomly divided into 4 groups, i.e. ischemia reperfusion injury group (IRI group), sulforaphane group (SFN group), LY294002+ ischemia reperfusion injury (LY+IRI group), and LY294002+ sulforaphane group (LY+SFN group). Isogeneic heterotopic heart transplantation model was established according to Li's method. Donor hearts storaged in histidine-tryptophan-ketoglutarate solution for 9 h were transplanted into the abdominal cavities of recipient rats to establish rat models of heterotopic heart transplantation. Akt, p-Akt, GSK-3β and p-GSK-3β protein expressions in the grafts were detected by immunohistochemistry and Western bolt 24 h after reperfusion. Results Compared with the IRI group, p-Akt and p-GSK-3β levels increased in the SFN group (P < 0.05). After using blocker LY294002, there was no significant difference in the expression of p-Akt or p-GSK-3β between the LY+SFN group and the LY+IRI group (P > 0.05). Conclusions SFN reduces cold ischemia reperfusion injury in heart transplantation of rats through activating Akt/ GSK-3β signaling pathway.