NR2A-Wnt-TLR2 signaling axis mediates neuronal mitochondrial dysfunction contributing to neuropathic pain induced by nerve injury in diabetic mice.

Diabetic neuropathy, a prevalent complication of diabetes mellitus (DM), induces neuropathic pain (NP) through spinal dorsal horn (SDH) central sensitization, with mitochondrial dysfunction emerging as a pivotal contributor. While N-methyl-d-aspartate receptor (NMDAR) subunit NR2A and Toll-like receptor 2 (TLR2) are implicated in NP pathogenesis, their mechanistic interplay in DM-associated mitochondrial dysregulation remains undefined. Here, we systematically investigated their involvement in central sensitization using gain-of-function and loss-of-function strategies for NR2A and TLR2. Firstly, a reduced mechanical paw withdrawal threshold (PWT) following spinal nerve ligation (SNL) in DM mice was found, which was also alleviated by NR2A and TLR2 knockout (KO), and reactive oxygen species (ROS) inhibition. Additionally, the NR2A-Wnt-TLR2-MyD88-nuclear factor kappa (NF-κB) signaling axis was activated in the SDH following SNL under DM. Interestingly, NP in DM background led to elevated ROS and reduced adenosine triphosphate (ATP), along with lower autophagy activity, as indicated by increased LC3-II and P62 and decreased autophagy-related genes. These mitochondrial injury-related changes were blocked by silencing TLR2 and rescued by its overexpression (oe). Furthermore, the alterations in ROS and ATP were also inhibited by autophagy activation. In conclusion, our findings elucidated a novel NR2A-Wnt-TLR2-NF-κB signaling axis that regulated mitochondrial dysfunction in the SDH after SNL under DM, contributing to accumulation of ROS, depletion of ATP, and impairment of autophagy. Targeting this signaling axis may have clinical potential in predicting and alleviating peripheral nerve damage in patients with DM.