Artesunate ameliorates acute pancreatitis by enhancing p62-mediated zymophagy through non-canonical NRF2 activation

Abstract Objective Zymophagy is a selective autophagy that eliminates activated trypsinogen during acute pancreatitis (AP). However, the upstream regulatory mechanisms and therapeutic potential of zymophagy remain poorly understood. This study investigated the mechanisms of zymophagy activation and its protective role in AP, using artesunate (AS) as a pharmacological tool. Methods AP models were induced by cerulein combined with lipopolysaccharide and evaluated by histopathological examination and enzymatic assays. Gene expression was knocked-down with specific shRNA or siRNA, and expression levels were detected by real-time PCR. Zymophagy and its regulatory mechanisms was characterized through multiple experiments, and molecular interactions were validated by co-localization, immunoprecipitation (IP), chromatin immunoprecipitation (ChIP), thermal shift assay, and molecular docking. Results AS mitigated AP in both in vivo and in vitro models by inhibiting trypsinogen hyperactivation in acinar cells. This protective effect was mediated by enhanced Atg5-mediated zymophagy activation via a p62-dependent and vacuole membrane protein 1 (VMP1)–ubiquitin-involved pathway, which promotes the targeted engulfment and degradation of trypsinogen. Mechanistically, AS binds the PB1 domain of p62, strengthens its interaction with Kelch-like ECH-associated protein 1 (Keap1), triggers ubiquitin–p62-mediated autophagic degradation of Keap1, facilitates the nuclear translocation of nuclear factor erythroid 2-related factor 2 (NRF2) and its selective binding to the antioxidant response element (ARE) in the p62 promoter, and ultimately upregulates p62 transcription. Conclusion Zymophagy is activated via a VMP1–ubiquitin–p62-dependent and Atg5-mediated canonical autophagy pathway. AS ameliorates AP by enhancing p62-mediated zymophagy via non-canonical NRF2 activation, which provides mechanistic evidence for therapeutic targeting of acinar zymophagy in AP management.