<p>Extracellular vesicles (EVs) from adipose-derived mesenchymal stem cells (ADSCs) may exert a therapeutic benefit in sepsis-associated organ dysfunction by delivering cargos to target cells. The aims of this study were to explore the therapeutic efficacy and potential mechanism of microRNA (miR)-574-5p delivered by ADSC-EVs in sepsis-associated acute kidney injury (SA-AKI). EVs were isolated from mouse ADSCs and characterized by multilineage differentiation potency, morphology, and surface markers. miR-574-5p expression in ADSC-EVs was analyzed by biochemical testing. Luciferase reporter and RNA pull-down assays were employed to identify the binding relation between miR-574-5p and glucose transporter 1 (GLUT1). Cecal ligation and puncture (CLP)-induced mouse SA-AKI model administered with ADSC-EVs was used to evaluate therapeutic effects on kidney injury. ADSC-EVs were co-cultured with RAW264.7 macrophages to assess effects on inflammation, glycolysis (lactate levels, extracellular acidification rate), and macrophage polarization. ADSC-EVs elevated miR-574-5p expression and polarized macrophages toward M2 phenotype, reducing kidney dysfunction and inflammatory response in CLP mice. In vitro, ADSC-EVs arrested lipopolysaccharide-induced M1 macrophage polarization and induced M2 macrophage polarization by upregulating miR-574-5p expression. Mechanistically, ADSC-EVs transferred miR-574-5p into macrophages, where miR-574-5p targeted GLUT1 and limited its expression. This effect contributed to the inhibition of glycolysis and promotion of M2 macrophage polarization. In vivo studies confirmed these positive effects of ADSC-EVs in SA-AKI via the miR-574-5p/GLUT1 pathway. Altogether, our findings suggest that ADSC-EVs carrying miR-574-5p target GLUT1 to restrain glycolysis and prime M2 macrophage polarization in a SA-AKI model, paving the way for establishing a therapeutic approach for SA-AKI.</p> Graphical abstract <p></p>