Aim: Neuropathic pain (NP) arises due to lesion or disease in somatosensory nervous system. Recent reports indicate the role of Withania somnifera (WS) in various inflammatory pain conditions. The objective of the present study was to explore the possible protective effect of WS against chronic constriction injury (CCI) induced NP in rats. Materials and Methods: CCI of sciatic nerve was performed in male Wistar rats. Various behavioral parameters (thermal hyperalgesia, cold allodynia) followed by biochemical parameters (lipid peroxidation, reduced glutathione, catalase and nitrite) were assessed in sciatic nerves. Drugs were administered consecutively for 21 days from the day of surgery. CCI to sciatic nerve significantly caused thermal hyperalgesia, cold allodynia and oxidative damage in the sciatic nerves when compared with naive and sham control. Results: Chronic administration of WS (100 and 200 mg/kg, p.o.) significantly attenuated hyperalgesia, cold allodynia and oxidative damage (as indicated by reduction in lipid peroxidation, nitrite concentration, restoration of reduced glutathione and catalase activity). Further, L-NAME (5 mg/kg, i.p.) pre-treatment with WS (100 mg/kg, p.o.) significantly potentiated the protective effect of WS which was significant when compared with their effect per se However, L-arginine (100 mg/kg, i.p.) pre-treatment with WS (100 mg/kg, p.o.) significantly reversed the protective effects of WS in sciatic nerve. Conclusion: Result of present study suggests that nitric oxide mechanism could be involved in the protective effect of WS against CCI induced behavior alterations and oxidative damage in rats.