Semaglutide ameliorates doxorubicin-induced cardiotoxicity by ameliorating BNIP3-mediated mitochondrial dysfunction. Doxorubicin, a potent chemotherapy drug, causes significant cardiotoxicity, limiting its use. Semaglutide, a GLP-1 receptor agonist, has shown potential in improving cardiac function and reducing doxorubicin-induced damage. This study demonstrates that semaglutide alleviates doxorubicin-induced cardiac dysfunction by reducing BNIP3 expression in mitochondria, thereby improving mitochondrial function and reducing cardiac injury. RNA sequencing identified BNIP3 as a key gene involved in the protective effect of semaglutide against doxorubicin-induced cardiotoxicity. Overexpression of BNIP3 in mice counteracted the protective effects of semaglutide, indicating its role in mediating the protective mechanism. In vitro experiments showed that semaglutide, through the PI3K/AKT pathway, reduces BNIP3 expression in mitochondria, improving mitochondrial function and reducing oxidative stress. The study also found that semaglutide mitigates doxorubicin-induced mitochondrial dysfunction by activating the PI3K/AKT pathway, which decreases BNIP3 expression. This pathway enhances mitochondrial function, reduces oxidative stress, and improves cardiac function. The findings suggest that semaglutide could be a potential therapy for reducing doxorubicin-induced acute cardiotoxicity. The study highlights the importance of the PI3K/AKT/BNIP3 pathway in protecting against doxorubicin-induced cardiac injury.Semaglutide ameliorates doxorubicin-induced cardiotoxicity by ameliorating BNIP3-mediated mitochondrial dysfunction. Doxorubicin, a potent chemotherapy drug, causes significant cardiotoxicity, limiting its use. Semaglutide, a GLP-1 receptor agonist, has shown potential in improving cardiac function and reducing doxorubicin-induced damage. This study demonstrates that semaglutide alleviates doxorubicin-induced cardiac dysfunction by reducing BNIP3 expression in mitochondria, thereby improving mitochondrial function and reducing cardiac injury. RNA sequencing identified BNIP3 as a key gene involved in the protective effect of semaglutide against doxorubicin-induced cardiotoxicity. Overexpression of BNIP3 in mice counteracted the protective effects of semaglutide, indicating its role in mediating the protective mechanism. In vitro experiments showed that semaglutide, through the PI3K/AKT pathway, reduces BNIP3 expression in mitochondria, improving mitochondrial function and reducing oxidative stress. The study also found that semaglutide mitigates doxorubicin-induced mitochondrial dysfunction by activating the PI3K/AKT pathway, which decreases BNIP3 expression. This pathway enhances mitochondrial function, reduces oxidative stress, and improves cardiac function. The findings suggest that semaglutide could be a potential therapy for reducing doxorubicin-induced acute cardiotoxicity. The study highlights the importance of the PI3K/AKT/BNIP3 pathway in protecting against doxorubicin-induced cardiac injury.