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. The gene discussed is CD86; the disease is metabolic dysfunction-associated steatotic liver disease.