Neuropathic pain is an intractable disease which in case opioids provide inadequate analgesia. It has been previously reported that voluntary wheel running prevented persistent hyperalgesia in sciatic nerve ligation (PSNL) mice. However, the mechanism of the preventive effect of the voluntary wheel running on persistent hyperalgesia was unknown. Since the anterior cingulate cortex (ACC) has been known to play an important role in chronic pain, we investigated factors associated with the attenuated hypersensitivity on voluntary wheel running in the ACC of the PSNL mice.A partial sciatic nerve ligation mouse was produced by ligating the sciatic nerve on the left side. The von frey test was used for pain assessment. RNA sequencing was performed using ACC tissue isolated from PSNL mice 2 weeks and 3 months after surgery.Mechanical hyperalgesia persisted for 63 days after sciatic nerve ligation. On the other hand, mechanical hyperalgesia decreased 14 days after the introduction of voluntary wheel running. The expression of 228 genes was upregulated in ACC 2 weeks after PSNL. Among these genes, 26 genes were included in the downregulated genes by the application of voluntary wheel running for 3 months. Gene Ontology analysis using DAVID detected 26 of these genes involved in neuronal differentiation (BTG anti-proliferation factor 2, CCAAT/enhancer binding protein beta, immediate early response 2, nuclear receptor subfamily 4, group A, member 2) were detected.These results suggest that the factor related to neuron differentiation in the ACC might be critical for attenuation of neuropathic pain.