Moreover, our present experiments demonstrated that miR-9-5p shuttled by hepatocytes-derived EVs induced inflammatory cell infiltration and inflammatory reaction, as evidenced by elevated levels of macrophage inflammatory factors, positive rates of CD86+ and CD11b+, and levels of macrophage surface markers (IL-1β, IL-6, and TNF-α), as well as levels of TG, TC, AST and ALT (in vivo) in the NAFLD model, thereby contributing to M1 macrophage polarization in NAFLD. This evidence concerns the gene ITGAM and metabolic dysfunction-associated steatotic liver disease.