Chronic nonspecific low back pain (CNLBP) is a leading cause of disability and remains a major burden for many public health systems. Acupuncture is a nonpharmacological treatment for CNLBP that can be effective in improving low back pain; nevertheless, its effect on improving back muscle endurance in patients with CNLBP and its duration of effect have not been studied. The goal of this study was to assess the impact of acupuncture on lower back muscle activity in CNLBP patients. This was a single-blind, randomized, crossover experimental study. Thirty female patients were randomized into Group A (15 patients) or Group B (15 patients). Patients in Group A were assigned to receive real acupuncture (RA) in the first phase and sham acupuncture (SA) in the second phase, while those in Group B received SA first and then RA, with a 1-week washout period between phases. Two-way repeated ANOVA was used to evaluate the effect of group and time on isokinetic parameters, Surface electromyography (sEMG) data, and blood data. Significant interaction effects were identified between group * time on the isokinetic parameters of the lumbar extensor muscles, sEMG values of the erector spinae, blood lactate levels, and blood ammonia levels (all p< 0.05). Compared with those of the SA group, the isokinetic parameters of the lumbar extensor muscles, sEMG values of the erector spinae, blood lactate levels, and blood ammonia levels of the RA group were significantly different (all p< 0.05). RA improves lumbar extensor endurance in patients with CNLBP and lasts approximately 9 minutes. RA can improve blood circulation to reduce blood lactic acid and blood ammonia produced during exercise.