The practice of acupuncture is becoming increasingly popular in veterinary medicine, especially as a method of providing pain relief. Originally based on principles derived from centuries of observation, conventional scientific mechanisms of action for acupuncture as a pain-relieving modality have recently been elucidated. Acupuncture points allow access to multiple regions of the body via the peripheral nervous system and its connection with the central nervous system. Local, segmental (spinal), and suprasegmental (brain) effects of acupuncture involve enhanced release of pain-relieving endogenous substances (e.g., opioids) and mitigated release of pain-inducing substances (e.g., inflammatory cytokines). In addition, there is evidence that acupuncture can induce positive neurochemical and cytoarchitectural change in the central nervous system via the phenomenon of neuroplasticity. Electroacupuncture is considered the most effective type of acupuncture delivery, allowing for more potent and long-lasting pain relief than is achieved via other methods (e.g., dry needling). The purpose of this review article is to summarize the relevant scientific literature from the last two decades relating to the physiological mechanisms of action of acupuncture as a pain-relieving modality.