Objective To investigate the protective effects and molecular mechanisms of growth differentiation factor 15 (GDF15) against doxorubicin-induced myocardial injury. Methods A doxorubicin-induced cardiomyopathy (DIC) model was established by intraperitoneal injection of doxorubicin (DOX, 15 mg/kg) in C57BL/6 mice. Forty SPF-grade 8-week-old male C57BL/6 mice were randomly divided into four groups: Con group, DIC group, DIC+rGDF15 group, and DIC+Ab-GDF15 group, with 10 mice in each group.Eighty SPF-grade 8-week-old male C57BL/6 mice were randomly divided into four groups:Con group, Con+rGDF15 group, DIC group, and DIC+rGDF15 group, with 20 mice in each group. The levels of serum myocardial injury markers, cardiac systolic function, myocardial oxidative stress and inflammation, as well as the apoptosis rate of cardiomyocytes, were evaluated in each group. Transcriptome sequencing was utilized to screen for potential molecular regulatory pathways. Results Compared with the Con group, the expression of GDF15 in the serum and myocardium of mice in the DIC group was increased (P＜0.01), the levels of serum myocardial injury markers were elevated (P＜0.01), and cardiac systolic function was impaired (P＜0.01). Treatment with rGDF15 significantly alleviated DOX-induced myocardial injury and cardiac dysfunction (P＜0.01), whereas treatment with Ab-GDF15 further exacerbated DOX-induced myocardial injury and cardiac dysfunction (P＜0.01). Additionally, compared with the Con group, the level of oxidative stress in the DIC group was elevated (P＜0.01), the inflammatory response was intensified (P＜0.01), the apoptosis rate of cardiomyocytes was increased (P＜0.01), and the AMPK signaling pathway was suppressed (P<0.01). Compared with the DIC group, the level of oxidative stress in the DIC+rGDF15 group was decreased (P＜0.01), the inflammatory response was alleviated (P＜0.01), the apoptosis rate of cardiomyocytes was reduced (P＜0.01), and the AMPK signaling pathway was upregulated (P＜0.01). Conclusion GDF15 exerts its protective effects by inhibiting oxidative stress and inflammatory response through the regulation of the AMPK signaling pathway, thereby alleviating doxorubicin-induced myocardial injury and cardiac dysfunction