Abstract: To investigate the therapeutic effects of ZLY16, a novel peroxisome proliferator-activated receptor (PPAR) β agonist, on Duchenne muscular dystrophy (DMD), C57BL/10ScSnJGpt-Dmdem3Cd4/Gpt (mdx) mice were gavaged with 30 mg/kg ZLY16 for 6 weeks. Expression of proteins associated with muscle regeneration, exercise ability, blood lipids content and skeletal muscle damage in mdx mice were investigated by behavioral experiments, histopathology, blood biochemical analysis, immunofluorescence and Western blot. A high-fat-induced myoblast differentiation inhibition model was established to examine lipid content and myoblast differentiation-related protein expression in myoblasts using Nile Red staining, immunofluorescence and Western blot. The results demonstrated that ZLY16 increased muscle grip strength, reduced triglyceride (TG) and total cholesterol (TC) levels, attenuated muscle fiber necrosis, fibrosis and inflammatory cell infiltration, and promoted muscle regeneration in mdx mice. ZLY16 promoted myoblast differentiation and myotube fusion in vitro by reducing lipid accumulation in murine skeletal muscle myoblast line (C2C12) cells. These findings suggest that ZLY16 improves motor function in mdx mice by decreasing lipid accumulation and promoting muscle regeneration.