ALYREF Promotes Pyroptosis in Severe Acute Pancreatitis by Inhibiting the Activation of ATK/mTOR Signaling Pathway Through m5C Modification on AKT1

Severe acute pancreatitis (SAP) is a digestive system disease with a high mortality rate. Acinar cell pyroptosis is crucial for the pathogenesis of SAP, but its mechanism has not been fully elucidated. m5C modification is involved in the progression of multiple diseases, but its role in SAP remains unclear. In this study, we aimed to investigate the mechanism of pyroptosis in SAP mediated by m5C modification. In vivo and in vitro SAP model were established by inducement of caerulein and cholecystokinin on C57BL/6 mice and 266‐6 cells. Hematoxylin eosin staining, TUNEL assay, ELISA, flow cytometry and western blot were performed to evaluate levels of inflammation and pyroptosis in vivo and in vitro of SAP. The underlying mechanism was detected by dot blot, methylated RNA Immunoprecipitation (MeRIP), RIP, dual luciferase reports and immunofluorescence staining. Results showed that inflammation and pyroptosis were increased in in vivo and in vitro SAP models, with the enhancement of m5C modification and ALYREF expression. Moreover, ALYREF knockdown inhibited inflammatory infiltration of pancreatic tissue and pyroptosis in the SAP mouse and cell models. ALYREF knockdown enhanced AKT1 expression by decreasing m5C modification on AKT1, and activated AKT/mTOR signaling pathway, while inhibiting the activation of AKT/mTOR pathway reversed the inhibition of pyroptosis induced by ALYREF knockdown. Collectively, we demonstrated that ALYREF knockdown inhibited pyroptosis in SAP through activating AKT/mTOR signaling pathway by decreasing m5C modification on AKT1. This study may provide a novel target and basis for the treatment of SAP.