Inhibition of PTP1B Through NADH Dehydrogenase Activity Reduces Acute Pancreatic Acinar Cell Pyroptosis by Inhibiting the ROS/TXNIP/NLRP3 Pathway.

OBJECTIVE

This study aims to investigate the role and regulatory mechanisms of PTP1B in acute pancreatitis.

METHODS

An in vitro AP cell model was established using Caerulin (CAE). Cells were subsequently treated with MSI-1436 (PTP1B inhibitor), ruscogenin (TXNIP inhibitor), N-acetyl-L-cysteine (NAC, ROS scavenger), and rotenone (mitochondrial complex I inhibitor), the levels of IL-1β and IL-18 in the cell supernatant were quantified using enzyme-linked immunosorbent assay (ELISA). Propidium iodide (PI) staining was visualized using fluorescence microscopy, and the fluorescence intensity of GSDMD was assessed via immunofluorescence. Western blotting was performed to detect the expression and activation of pro-caspase-1, cleaved-caspase-1, cleaved-caspase-4/5/11, AP-1, ASC, and NLRP3. Mitochondrial membrane potential was assessed using the JC-1 assay, complex I activity was determined with the MitoCheck™ Complex I Activity Assay Kit, and ROS levels were measured using a ROS detection kit.

RESULTS

CAE treatment significantly increased the secretion of inflammatory cytokines IL-1β and IL-18 in pancreatic acinar cells. Expression levels of PTP1B, pyroptosis-related factors-including AP-1, ASC, cleaved-Caspase4, cleaved-Caspase5, cleaved-Caspase11, cleaved-Caspase1, and NLRP3-were significantly upregulated. However, inhibition of PTP1B and TXNIP markedly reduced inflammation and pyroptosis, suggesting that the PTP1B/TXNIP axis mediates CAE-induced cellular injury. Furthermore, suppression of ROS generation and mitochondrial complex I activity also attenuated CAE-induced inflammation and pyroptosis in pancreatic acinar cells.

CONCLUSION

Targeting PTP1B mitigates CAE-induced pyroptosis in pancreatic acinar cells by restoring NADH dehydrogenase activity and suppressing ROS/TXNIP/NLRP3 inflammasome activation.