Spinal neuronal ferroptosis aggravates peripheral nerve injury-induced neuropathic pain via modulating competing endogenous RNA axis - RNA-Seq
Peripheral nerve injury-induced neuropathic pain presents a significant clinical challenge and the pathogenesis remains unclear. Recent findings have implicated spinal neuronal ferroptosis in the modulation of neuropathic pain progression, but the molecular mechanism is inadequately understood. This work constructed competitive endogenous RNA (ceRNA) regulatory network in neuronal ferroptosis model through total transcriptome sequencing. Bioinformatic analyses elucidated the biological roles and signaling pathways of differentially expressed genes associated with neuronal ferroptosis. In vitro experiments substantiated the involvement of two critical regulatory axes, namely lncRNA-Sh3bp5l/miR-92b-5p/Pgd and lncRNA-Scrn2/miR-133a-3p/Srxn1, in the regulation of neuronal ferroptosis. The knockdown of PGD and SRXN1 was observed to significantly inhibit the ferroptosis phenotype. Utilizing an in vivo chronic constriction injury (CCI) pain model, we demonstrated that the expression of ceRNA-mediated downstream targets, PGD and SRXN1, was localized within spinal neurons.  Furthermore, lncRNA-Sh3bp5l and lncRNA-Scrn2 were found to be upregulated in the ipsilateral spinal cord following CCI injury, accompanied by increased levels of PGD and SRXN1. In brief, this study explored the key ceRNA axis that regulates spinal neuronal ferroptosis, which might serve as the potential therapeutic targets for neuropathic pain. Overall design: RNA sequencing of retinoic acid-induced N2a cells with or without RSL3 stimulation