Procaine alleviates microglial pyroptosis by inhibiting O-GlcNAcylation of NOD-like receptor family, pyrin domain-containing 3 through the Janus kinase 2/signal transducer and activator of transcription 3 signaling pathway.

BACKGROUND

Neuropathic pain is a chronic condition involving microglial pyroptosis mediated by the NOD-like receptor family, pyrin domain-containing 3 (NLRP3) inflammasome. Current treatments are limited, prompting the need for new therapies targeting these mechanisms. This study evaluates Procaine's effects on microglial pyroptosis and its underlying pathways.

METHODS

BV-2 cells were exposed to lipopolysaccharide (LPS) to induce pyroptosis. NLRP3 O-GlcNAcylation was assessed using wheat germ agglutinin pull-down and co-immunoprecipitation assays. ELISA was employed to measure interleukin (IL)-1β and IL-18 secretion levels. The transcriptional regulation of O-GlcNAc transferase (OGT) by signal transducer and activator of transcription 3 (STAT3) was investigated through dual-luciferase reporter and chromatin immunoprecipitation assays.

RESULTS

Procaine treatment markedly inhibited LPS-induced pyroptosis in BV-2 cells while promoting the viability. NLRP3 O-GlcNAcylation contributed to LPS-induced microglial pyroptosis. Mechanistically, the Janus kinase 2 (JAK2)/STAT3 signaling pathway promoted LPS-induced microglial pyroptosis by transcriptionally activating OGT expression. In addition, procaine inhibited LPS-induced microglial pyroptosis by repressing OGT-mediated NLRP3 O-GlcNAcylation through inactivating the JAK2/STAT3 pathway.

CONCLUSION

Procaine alleviated LPS-induced microglial pyroptosis by inhibiting OGT-mediated O-GlcNAcylation of NLRP3 through inactivating the JAK2/STAT3 signaling pathway. Our research provides a potential therapeutic strategy for neuropathic pain.