This study investigates the role of histone lactylation in sepsis-associated acute kidney injury (SA-AKI). The research reveals that histone H3K18 lactylation (H3K18la) is elevated in SA-AKI, particularly at the promoter region of RhoA, and is positively correlated with transcription. Inhibition of lactate levels reversed abnormal histone lactylation at the RhoA promoter. The study further demonstrates that histone lactylation promotes the RhoA/Rho-associated protein kinase (ROCK)/Ezrin signaling pathway, leading to inflammation, cell apoptosis, and renal dysfunction. Ezrin, a protein with multiple lactylation sites, is identified as a substrate for lactylation, with K263 being the primary site. Inhibition of GLUT1 and mutation of the K263 site reduced inflammation and apoptosis, suggesting a potential therapeutic target for delaying renal dysfunction in SA-AKI. The findings provide new insights into the epigenetic regulation of SA-AKI and highlight the potential of lactylation as a therapeutic target.This study investigates the role of histone lactylation in sepsis-associated acute kidney injury (SA-AKI). The research reveals that histone H3K18 lactylation (H3K18la) is elevated in SA-AKI, particularly at the promoter region of RhoA, and is positively correlated with transcription. Inhibition of lactate levels reversed abnormal histone lactylation at the RhoA promoter. The study further demonstrates that histone lactylation promotes the RhoA/Rho-associated protein kinase (ROCK)/Ezrin signaling pathway, leading to inflammation, cell apoptosis, and renal dysfunction. Ezrin, a protein with multiple lactylation sites, is identified as a substrate for lactylation, with K263 being the primary site. Inhibition of GLUT1 and mutation of the K263 site reduced inflammation and apoptosis, suggesting a potential therapeutic target for delaying renal dysfunction in SA-AKI. The findings provide new insights into the epigenetic regulation of SA-AKI and highlight the potential of lactylation as a therapeutic target.