Abstract In non-diabetic mice, s.c. injection of formalin to the hindpaw had a biphasic effect: an immediate nociceptive response (first-phase) followed by a tonic response (second-phase). However, only the immediate nociceptive response was observed in diabetic mice. The duration of the first-phase response was significantly longer in diabetic mice than in non-diabetic mice. In diabetic mice, when spantide, an antagonist of substance P, reduced the duration of the nociceptive response in the first-phase to the levels that were observed in non-diabetic mice, the second-phase response appeared. The second phase also became apparent in diabetic mice after pretreatment with naltrindole (3 mg/kg), an antagonist of δ-opioid receptors. These results suggest that a negative control system, which is mediated by δ-opioid receptors and links substance P with somatostatin-mediated nociceptive transmission, may inhibit the formalin-induced second-phase of the nociceptive response in diabetic mice.