Molecular changes caused by metabolic reprogramming in ischemia and reperfusion cortex remain an unaddressed issue. Applying the quantitative proteomic and lactylproteomic analysis, we found that the molecular changes of ischemic brain tissues are quite different between ischemia and reperfusion stages. Conjoint analysis with a previous published transcriptome data further showed that the significant changes in protein levels and protein lactylations were mainly correlated with neurons in the reperfusion and ischemia stages, respectively. The significant upregulated lactylation of Aldoc, one of the glycolytic enzymes, at lysine 230 may contribute to the metabolic reprogramming by regulating glycolysis of astrocyte in ischemia phase, and which may further promote the lactate-derived lactylation of protein lysine residues. Our findings lay the foundation for future mechanistic studies of metabolic reprogramming from a novel perspective of lactylation, and illustrate that protein lactylations may play important roles in regulating the biological functions of neural cells after ischemic stroke.