In this study, utilizing both in vivo (LAD-operated mice) and in vitro (hypoxic AC16 cardiomyocytes) models, we have identified PFKP-K688 site lactylation (PFKP-K688la) as a crucial metabolic regulatory target in myocardial hypoxia. Through lactated proteomics analysis, we detected 521 Kla modified proteins, with PFKP emerging as the principal target of Kla modification.