Comprehensive profiling and molecular characterization of alternative splicing regulation in synaptic remodeling associated with neuropathic pain induced by chronic constrictive injury in a rat Model

BACKGROUND: Neuropathic pain (NP) is a chronic pain condition caused by injury to the nervous system. Although alternative splicing dysregulation has been implicated in NP, the contribution of RNA-binding proteins (RBPs) to this process remains unclear. METHODS: We analysed 12 RNA-seq datasets (GSE217932) using the SUVA pipeline to identify splicing alterations in Sham, chronic constriction injury (CCI), and CCI treated with L-tetrahydropalmatine (CCI_THP) groups. Co-expression and co-disturbance analyses were performed to define splicing-associated RBP networks. Key findings were further validated in a CCI rat model by behavioural testing, junction-specific RT-qPCR, immunohistochemistry, and immunofluorescence. RESULTS: , were significantly upregulated in CCI. These results were supported by in vivo validation of selected splicing events and protein changes. CONCLUSIONS: This study provides a transcriptome-wide view of alternative splicing and RBP dysregulation in NP, and suggests that THP may alleviate neuropathic pain partly by modulating aberrant splicing programs. These findings offer insight into NP mechanisms and identify potential therapeutic targets.