Histone H3K18 lactylation and Ezrin lactylation are significantly elevated in sepsis-associated acute kidney injury (SA-AKI), contributing to renal dysfunction. This study reveals that increased lactate levels in SA-AKI promote histone lactylation, particularly at the promoter of RhoA, leading to RhoA/ROCK/Ezrin signaling activation, inflammation, and apoptosis. Ezrin undergoes lactylation at multiple sites, with K263 being the primary site. Inhibition of lactate metabolism or Ezrin K263 lactylation reduces inflammation and apoptosis, suggesting these modifications as potential therapeutic targets. The findings highlight the role of histone lactylation in SA-AKI pathogenesis and provide insights into the epigenetic regulation of kidney injury. The study also identifies the involvement of Ezrin in glycolytic reprogramming and GLUT1 sublocalization, further linking lactylation to renal dysfunction. Overall, the results suggest that targeting lactylation pathways may offer new strategies for improving renal function in SA-AKI.Histone H3K18 lactylation and Ezrin lactylation are significantly elevated in sepsis-associated acute kidney injury (SA-AKI), contributing to renal dysfunction. This study reveals that increased lactate levels in SA-AKI promote histone lactylation, particularly at the promoter of RhoA, leading to RhoA/ROCK/Ezrin signaling activation, inflammation, and apoptosis. Ezrin undergoes lactylation at multiple sites, with K263 being the primary site. Inhibition of lactate metabolism or Ezrin K263 lactylation reduces inflammation and apoptosis, suggesting these modifications as potential therapeutic targets. The findings highlight the role of histone lactylation in SA-AKI pathogenesis and provide insights into the epigenetic regulation of kidney injury. The study also identifies the involvement of Ezrin in glycolytic reprogramming and GLUT1 sublocalization, further linking lactylation to renal dysfunction. Overall, the results suggest that targeting lactylation pathways may offer new strategies for improving renal function in SA-AKI.