Objectives: This study was conducted to investigate the protective effect and intervention mechanism of C1 (a selective AMPK activator) on muscle damage in mice with Duchenne muscular dystrophy (DMD). Methods: We administered C1 (18 mg/kg and 54 mg/kg) to 10-week-old male DMD mice for 8 weeks. A series of behavioral tests were carried out to assess motor ability. Skeletal muscle samples were subjected to pathological analysis, western blot and proteomic analysis to explore the different protein networks associated with muscle pathology. Compound C, an AMPKα inhibitor, has been used as a negative control for follow-up experiments to verify the efficiency of C1 on AMPK activation. Key findings： C1 treatment significantly enhanced motor function and reduced muscle fibrosis and inflammation in DMD mice. We then found that C1 activated the AMPKα/PGC-1α signaling pathway, increasing ATP levels and mitochondrial DNA copy number in DMD mice. These improved effects on mitochondrial function were reversed by Compound C. In addition, Gastrocnemius muscle proteomics showed that C1 attenuated oxidative stress damage and improved muscle structural development. Conclusion: Collectively, C1 alleviates muscle damage in DMD mice by activating AMPKα to improve mitochondria.