Danshenol B alleviates central post-stroke pain by regulating PIK3CG/NLRP3 signaling pathway
Background: Central post-stroke pain (CPSP) severely impairs quality of life and is challenging to manage, potentially leading to self-harm or suicide. Although Compound Salvia miltiorrhiza (Danshen) is effective against neuropathic pain, its specific active components and mechanisms remain unclear. Study design and methods: We established a CPSP mouse model and performed RNA sequencing on the ipsilateral Ventral posterolateral thalamic nucleus and Ventral posteromedial thalamic nucleus (VPL/VPM) to identify differentially expressed genes (DEGs). DEGs were analyzed through network pharmacology and molecular docking to assess interactions with 42 Danshen components. We evaluated the effects of varying Danshenol B concentrations, administered daily into the VPL/VPM, on CPSP. Changes in PIK3CG and NLRP3 were monitored to determine Danshenol B's analgesic effects. Additionally, we overexpressed and knocked down PIK3CG to examine its role in CPSP and assessed NLRP3 to explore if it acts downstream of PIK3CG. Results: In the CPSP model, 409 DEGs were identified in the VPL/VPM, involved in pathways like signal transduction and NOD-like receptor signaling. Network pharmacology linked 21 Danshen components to 11 DEGs. Molecular docking showed strong binding affinities (=?-7.00?kcal/mol) between four Danshen components—tanshinaldehyde, sclareol, MOL007155, and Danshenol B—and proteins including BTK, CASP1, MMP9, and PIK3CG. Danshenol B (0.5?µM) significantly alleviated CPSP and reduced PIK3CG and NLRP3 protein. PIK3CG overexpression increased NLRP3 and counteracted Danshenol B's effects, while its knockdown induced CPSP and reduced NLRP3 expression. Conclusion: Danshenol B alleviates CPSP by suppressing the PIK3CG/NLRP3 signaling pathway, clarifying its therapeutic mechanism, and highlighting its potential in analgesic drug development. Overall design: To investigate differentially expressed genes in the CPSP model, we established this model by injecting 0.01 U/10 nL of type IV collagenase into the right VPM/VPL of C57 mice; controls received 10 nL saline. Fourteen days post-injection, VPM/VPL tissues were harvested for transcriptomic sequencing to identify DEGs. We then predicted Danshen components and target genes, analyzing their interactions with DEGs and evaluating their therapeutic effects on CPSP.