Data-independent acquisition-based quantitative proteomic analysis of the effects of environmental enrichment on the dorsal roots of mice with neuropathic pain induced by chronic sciatic nerve constriction injury.

Neuropathic pain (NP) significantly impairs patients' quality of life. Environmental enrichment (EE), a novel nonpharmacological therapy, enhances cognitive and behavioral outcomes by providing a structured and stimulating environment. Using data-independent acquisition (DIA)-based proteomics analysis, we aimed to screen out differential proteins that are implicated in the mechanisms by which EE ameliorates NP. Mice were exposed to either a standard environment (SE) or EE for 2 h daily over a 14-day pretreatment, followed by another 14-day treatment phase under the same conditions after chronic sciatic nerve constriction injury (CCI) to induce NP. The mechanical withdrawal threshold (MWT) and withdrawal thermal latency (WTL) were assessed on the day before CCI and on Days 3, 6, 9, 12, and 15 following CCI. On Day 6 post-CCI, spinal cord samples were collected for histopathology and apoptosis analysis, while lumbar dorsal roots were analyzed via DIA proteomics. Western blotting validated the findings. Our results indicated that EE reversed the decrease in the MWT, WTL, and inhibited apoptosis in CCI mice. A total of 5724 proteins were identified via DIA proteomics analysis, 262 of which may relate to the mechanism by which EE alleviates NP. Changes in mitochondrial function may be the primary mechanism of the therapeutic effect of EE. Western blot analysis revealed that EE may alleviate NP by downregulating the protein expression of TIMM22, RBBP4, and STOM and increasing the protein expression of MRPS28. In conclusion, EE seems to be an effective nonpharmacological approach for the treatment of NP, with mechanisms closely linked to mitochondrial function.