Chronic neuropathic pain was an intractable clinical problem and challenge to the quality of life of patients. It is essential to unveil the mechanisms that drive chronic pain so as to formulate new strategies for therapy of chronic pain. The medial prefrontal context (mPFC) plays a pivotal role in pathogenesis of chronic pain. However, its underlying molecular mechanisms are largely elusive. Herein, we demonstrated that mitogen-activated protein kinase-phosphatase1 (MKP1), a negative regulatory factor of mitogen activated protein kinases (MAPKs), was activated and persistently upregulated in the mPFC neurons by qPCR and western blotting assays following chronic constrictive injury (CCI) in mice. Inhibition of MKP1 in the mPFC contralateral to the injury site could reverse CCI-induced pain behavior and neuronal activity either via employment of BCI (MKP1 antagonist) or Lenti-MKP1 particles. Furthermore, we identified the substrates of MKP1 in the mPFC involved in chronic neuropathic pain. The western blot results showed that the phosphorylation of p38 and JNK1/2 was significantly decreased after CCI, while the phosphorylation of ERK1/2 was significantly increased, suggesting that p38 and JNK1/2 were the substrates of MKP1 in mouse mPFC, but not ERK1/2. Additionally, microinjection of BCI in the mPFC contralateral to the injury side could reverse downregulation of p- p38 and p- JNK1/2 in CCI mice. SB203580 (p38 inhibitor) or SP600125 (JNK1/2 inhibitor) could reverse BCI-induced analgesia. Our findings validated that MKP1 in the mPFC modulated chronic neuropathic pain via p38 and JNK1/2, suggesting a possible MKP1-mediated process would participate in neuronal transmission pathways implicated in pain modulation.