Objective Exploring the influences of intraperitoneal injection of dexmedetomidine(Dex) on activation of spinal glial cells and expression of inflammatory factors in rats with diabetic neuropathic pain(DNP). Methods Thirty two health male SD rats, weight 180-200 g, were divided into four groups (n=8) according the random number table method. The four groups included the control group(group C), diabetic neuropathic pain group(group DNP), group Dex and α2 receptor antagonist group(group YOH). Streptozotocin(STZ) was used to establish the DNP model in rats. Starting from 28 d after STZ injection, Dex group was given intraperitoneal injection of Dex(50 μg/kg) for consecutive 7 d, while group YOH was given intraperitoneal injection of Yohimbine(0.1 mg/kg) 30 min before intraperitoneal injection of Dex(50 μg/kg) for consecutive 7 d. Mechanical withdrawal threshold (MWT) was determined in each group of rats 29-35 d after STZ injection. Rats were sacrificed after MWT detection on 35 d. Immunofluorescence double-staining was used to detect activation of microglia and astrocytes in rat spinal cord of each group. ELISA was used to measure TNF-α and IL-1 expression in rat spinal cord of each group. Results Compared with group C, rat body weights were decreased while blood glucose values were increased in group DNP, group Dex and group YOH(P 0.05). Compared with group C, MWT at all time points decreased in group DNP and group YOH(P 0.05). Conclusions Dex can relieve hyperalgesia in rats with diabetic neuropathy pain. The underlying mechanism may be related to inhibition of microglia activation and reduction of the inflammatory reaction in spinal cord. Key words: Dexmedetomidine; Diabetic neuropathic pain; Microglia; Astrocyte; Inflammatory reaction