Objective To investigate the effects of electroacupuncture on NF-κB p65 expression in the spinal cord dorsal horn of rats with diabetic neuropathic pain. Methods Thirty SD rats were randomly divided into the normal and model groups. The rats in the model group were fed a high-fat and high-sugar diet for 5 weeks, then intraperitoneally injected with streptozotocin (STZ). The model diabetic neuropathic pain rats were further randomly divided into the diabetic neuropathic pain (DNP) model group and the diabetic neuropathic pain model + EA (DNP + EA) group. Two hertz of electroacupuncture was administered at the Zusanli and Kunlun ipsilateral acupoints for 7 consecutive days. Changes occurred in the fasting blood glucose, paw withdrawal threshold and paw withdrawal latency. Hematoxylin-eosin ( HE) staining was used to observe the morphological changes in the sciatic nerve in the rats, and western blotting was used to detect the NF-κB p65 protein expression in the spinal cord dorsal horn in rats. Results After 5 weeks on the high-fat and high-sugar diet followed by a single intraperitoneal injection of STZ, the fasting blood glucose increased, and the paw withdrawal threshold and latency decreased, indicating that the DNP model was successfully established. Electroacupuncture increased the paw withdrawal threshold and latency in the DNP rats but did not significantly affect the fasting blood glucose. HE staining result showed disordered arrangement of the myelinated nerve fibers, axonal swelling, uneven density of the myelin sheath and vacuolar degeneration of the sciatic nerve in the DNP rats. Electroacupuncture intervention did not significantly affect the sciatic nerve morphology in the DNP rats. Western blotting showed significantly increased NF-κB p65 protein expression in the spinal cord dorsal horn of the DNP rats, and electroacupuncture downregulated the NF-κB p65 protein expression in the spinal cord dorsal horn of the DNP rats. Conclusions Low frequency electroacupuncture has a good analgesic effect on the DNP model rats, possibly related to its inhibiting NF-κB p65 expression in the spinal cord dorsal horn.