Objective To investigate the effect of reactive oxygen species(ROS) on P2X3 receptor-mediated neuropathic pain. Methods Neuropathic pain model was induced in female Sprague Dawley rats by chronic compression of dorsal root ganglia (CCD), and the CCD rats were randomly divided into 4 groups (each group containing 10 rats): Saline group (NS group); PBN 100 mg/kg treatment group; PBN 30 mg/kg treatment group; and PBN 10 mg/kg treatment group. The specific agonist of P2X3 receptor, α, β-meATP (50 nmol in 50 μL), was subcutaneously injected into the plantar surface of the right hind paws of each rat 30 min after PBN or NS injection. The spontaneous paw flinching times and withdrawing time were observed for 15 min after injection and the paw lift time per minute (PLTPM) in every 2 minutes was calculated. Results Pre-treatment with PBN inhibited α, β-meATP-induced spontaneous pain in a dose-dependent manner. Compared with the NS group, PBN 100 mg/kg group significantly inhibited flinching response during the first 6 min (P<0.05), while the rats in PBN 30 mg/kg group only had significantly attenuated flinching response during the second to the fourth minute compared with NS group(P<0.05). Conclusion Oxygen free radical scavenger can effectively alleviate the neuropathic pain caused by CCD and P2X3 receptor agonist-induced spontaneous pain. ROS may act as a messenger in P2X3 receptor-mediated pain signaling transmission.