Antiretroviral therapy (ART) often results in painful peripheral neuropathy. Given that voluntary exercise has been shown to be beneficial in terms of modulating pain-like behaviors in various animal models of peripheral neuropathy, we have investigated the effects of voluntary wheel running on neuropathic pain induced by chronic ART. We first established an animal model of peripheral neuropathy induced by chronic 2',3'-dideoxycytidine (ddC) treatment. We showed that mice receiving ddC (3 mg/kg/day) had increased mechanical and thermal sensitivity at 9 weeks after the onset of the treatment. We also found that voluntary wheel running attenuated or delayed the onset of ddC-induced peripheral neuropathy. This phenomenon was associated with the attenuation of dorsal root ganglion nociceptive neuron membrane excitability and reduction in the expression of the transient receptor potential cation channel subfamily V member 1 (TRPV1). Taken together, these results suggest that voluntary exercise is an effective strategy by which ART-induced peripheral neuropathy can be alleviated.