Effect of Esketamine Pretreatment on Necroptosis in Mice with Trigeminal Neuralgia and Anxiety-Like Behavior via the RIP1/RIP3/MLKL Pathway

To explore the effect of esketamine (ES) pretreatment on necroptosis in mice with trigeminal neuralgia accompanied by anxiety-like behavior and its correlation with the RIP1/RIP3/MLKL signaling pathway. Thirty male C57BL/6J mice were randomized into five groups ( n =6): Control, Sham, model group (PIONT), PIONT+NS, and PIONT+ES. Trigeminal neuralgia model was established by left infraorbital nerve transection. ES (10 mg/kg, i.p.) was given daily for 5 days preoperatively (preoperative pretreatment) in the PIONT+ES group before molding, and equal-volume saline was administered identically in the PIONT+NS group. Pain thresholds were measured on postoperative days 3, 7, 14 and 21. Behavioral assessments were conducted on day 19, and tissue samples were harvested on day 21. Histopathological staining, immunofluorescence, qPCR, Western blot, ELISA and molecular docking were further performed for subsequent analyses. PIONT group showed reduced pain thresholds, anxiety-like behaviors, neuronal degeneration, decreased dendritic spine density, elevated TNF-α, and upregulated RIP1/RIP3/MLKL (all P <0.05). ES pretreatment prevented the development of these pathological alterations, with improved neuropathology, reduced TNF-α/RIP1/RIP3/MLKL levels, and increased spine density ( P <0.05). Molecular docking showed ES binds RIP1/RIP3/MLKL/TNF-α with energies -5.46, -5.10, -5.94, -7.36 kcal/mol. ES pretreatment prevents the development of trigeminal neuralgia and anxiety-like behaviors in mice, which is associated with suppression of necroptosis-related signaling.