BACKGROUNDS: Takotsubo cardiomyopathy (TTC), also known as stress-induced cardiomyopathy, is a condition characterized by transient left ventricular dysfunction without coronary artery obstruction. METHODS: We utilized label-free quantitative proteomics to analyze protein expression in a murine model of TTC, induced by a high dose of isoproterenol (ISO) injection. RESULTS: We found that a single high dose of ISO injection in mice could induce stress-related cardiac dysfunction.The proteomic analysis revealed 81 differentially expressed proteins (DEPs) between the ISO and control groups-39 were upregulated, and 42 were downregulated. Key pathways enriched by Gene Ontology (GO) analysis included collagen fibril organization, cholesterol biosynthesis, and elastic fiber assembly. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment indicated significant changes in unsaturated fatty acid biosynthesis, glutathione metabolism, steroid biosynthesis, and ferroptosis. Key hub proteins identified by the protein-protein interaction (PPI) network included Ntrk2, Fdft1, Serpine1, and Cyp1a1. Gene set enrichment analysis (GSEA) showed upregulation in terpenoid backbone biosynthesis, oxidative phosphorylation, and ferroptosis, with downregulation in pathways such as systemic lupus erythematosus and Rap1 signaling. CONCLUSIONS: This study employed high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify key proteins associated with energy metabolism, oxidative stress, inflammation, and cell death in TTC. These findings provide new insights into the molecular mechanisms of stress-induced myocardial injury and may offer potential therapeutic targets for mitigating cardiovascular damage under stress conditions.