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Neural sensitization can cause neuroinflammation, which is a type of inflammation that occurs in both the peripheral nervous system and central nervous system. The purpose of this study was to investigate the effect of dimethyl itaconate (DMI) on the expression of NGFI-A and NGFI-B and inflammatory cytokines in the spinal cord in the formalin test. The rats were divided into five groups: control, formalin, DMI 10 mg/kg + formalin, DMI 20 mg/kg + formalin and diclofenac sodium 10 mg/kg + formalin. We evaluated the impact of DMI on the spinal cords NGFI-A and NGFI-B expressions and inflammatory and anti-inflammatory cytokines [interleukin-1 beta (IL-1β), tumour necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10)]. The findings indicate that DMI 10, DMI 20 and diclofenac sodium 10 mg/kg can relieve pain in rats during the formalin test. In addition, these substances were found to reduce the expression of NGFI-A and NGFI-B in the spinal cord. Moreover, DMI 10, DMI 20 and diclofenac sodium 10 mg/kg were observed to increase the expression of IL-10 while decreasing IL-1β, TNF-α and IL-6 in the spinal cord when compared with the formalin group. We have found that administering DMI can alleviate pain in rats during formalin test. Through our research, we have observed that DMI decreases the expression of NGFI-A and NGFI-B in the spinal cord. Furthermore, DMI has been shown to increase the levels of IL-10 while decreasing IL-1β, TNF-α and IL-6 in the spinal cord.