Myopia has become the major cause of visual impairment worldwide. Although the pathogenesis of myopia remains controversial, proteomics studies suggest that dysregulation of retinal metabolism is potentially involved in the pathology of myopia. Lysine lactylation of proteins plays a key role in regulating cellular metabolism, but little is known about its role in the form-deprived myopic retina. In this study, we performed lactylation proteomic analysis of the retinas of form-deprived myopic guinea pigs and found downregulated levels of lactylation of metabolism-critical enzymes in the retina. As the first report on retinal lactylation in myopic eyes, this study provides a reliable basis for further studies on retinal lactylation in myopic eyes.