The study investigated the protective effects and underlying mechanisms of atractylone (Atr) against H₂O₂-induced oxidative damage in rat cardiomyocyte H9c2 cells. Initially, an oxidative damage model was established by inducing H9c2 cells with H₂O₂. The effects of Atr against oxidative damage were assessed by measuring cell viability, apoptosis, and oxidative stress markers. Bioinformatics analysis was employed to predict the molecular mechanisms of Atr. Further experiments were conducted to examine the impact of Atr on the phosphatidylinositol 3-kinase (PI3K)/ protein kinase B (AKT) signaling pathway and to verify its involvement in the regulation of oxidative damage in H9c2 cells. Results showed that Atr significantly alleviated oxidative damage in H9c2 cells (P<0.05). The PI3K/AKT signaling pathway appears to be involved in the regulation of Atr's protective effects. Atr significantly upregulated the suppressed PI3K/AKT signaling pathway in damaged cells (P<0.05), and its protective effects were reversed by pathway inhibitor (P<0.05). In conclusion, Atr exerts its protective effects against H₂O₂-induced oxidative damage in H9c2 cells by activating the PI3K/AKT signaling pathway.