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Introduction Stimulating endogenous cardiomyocyte proliferation is a promising therapeutic strategy for myocardial infarction (MI). 4’-O-Methylochnaflavone (4’-MF), a natural compound, has uncharacterized cardioprotective potential. This study aims to investigate whether 4’-MF promotes cardiac regeneration and to decipher its specific molecular mechanism.Methods In vivo, an adult mouse MI model was established. Cardiac function (echocardiography) and fibrosis (Masson staining) were evaluated. In vitro, proliferation markers (EdU, pH3, Ki67) were assessed in primary cardiomyocytes. Network pharmacology intersected potential targets of 4’-MF with MI-related genes. The key target (AKT) and PI3K-Akt signaling were prioritized through molecular docking and CETSA. Gain- (4’-MF) and loss-of-function (PI3K/Akt inhibitor LY294002; constitutively active GSK-3β mutant) experiments were performed.Results 4’-MF treatment significantly enhanced cardiomyocyte proliferation in vitro and improved cardiac function while reducing fibrosis in vivo post-MI. Mechanistically, 4’-MF directly bound and activated AKT, which specifically phosphorylated and inhibited GSK-3β, leading to β-catenin nuclear accumulation and upregulation of cell-cycle genes, Cyclin D1. Crucially, both pharmacological PI3K/Akt inhibition and genetic GSK-3β activation completely abolished 4’-MF-induced β-catenin signaling and cardiomyocyte proliferation, establishing a direct AKT-GSK-3β/β-catenin axis.Conclusion This study demonstrates that 4’-O-Methylochnaflavone promotes myocardial regeneration by directly targeting AKT to orchestrate a specific GSK-3β/β-catenin transcriptional program. This work moves beyond general pathway activation, defining a precise molecular cascade for natural product-induced cardiac repair.