Objective To determine the effect of the extracts of Ginkgo biloba leaf (EGB) on the endoplasmic reticulum stress-induced cardiomyocyte injury and investigate the role of Wnt signal pathway in this process. Methods Tunicamycin(Tm) was used to establish the endoplasmic reticulum stress model in rat cardiomyocytes which were isolated from neonatal rats in 1d after born and then cultured for 48h. There were 4 groups of cardiomyocytes, that is, control, Tm treated, Tm+EGB treated and EGB treated. MTT assay was used to detect cell viability. The dual-luciferase report system was used to measure Wnt activity. C-myc and CyclinD1 were detected by real-time PCR. Results Compared to control cells, Tm treatment resulted in significantly decreased cell viability, but the presence of EGB markedly attenuated the cell injury. The treatment also decreased the activity of Wnt, whereas co-treatment of Tm and EGB led to not only the increase in Wnt activity, but also recovery of the C-myc and CyclinD1 levels. However, Wnt inhibitor, secreted frizzled-related protein (sFRP) decreased Wnt activity, C-myc and CyclinD1 levels, and inversed EGB-induced protective effect. Conclusion These findings demonstrate that EGB protects cardiomyocytes against Tm-induced injury through improving Wnt activity.