Curcumol alleviates acute pancreatitis by inhibiting RIPK1/RIPK3/MLKL pathway-mediated necroptosis: integrated bioinformatics, network pharmacology, molecular docking and dynamics simulations, and experimental validation.

Abstract

BACKGROUND: Acute pancreatitis (AP) is a critical disease characterized by pancreatic tissue necrosis, a process primarily mediated by cellular necroptosis, yet safe and effective therapeutic agents are still lacking. Curcumol (CCM) is a bioactive compound extracted from plants of the Curcuma genus with anti-inflammatory and antioxidant properties; however, its role in AP remains unclear. OBJECTIVE: This study aimed to investigate the potential therapeutic effects of CCM in AP and elucidate whether CCM can alleviate pancreatic acinar cell injury by modulating necroptosis, particularly via the RIPK1/RIPK3/MLKL pathway. METHODS: This study integrated bioinformatics analysis, network pharmacology, molecular docking, molecular dynamics (MD) simulations, cellular thermal shift assay (CETSA), and experimental validation in AP models to investigate the potential effects of CCM in AP and its association with necroptosis. RESULTS: Results from bioinformatics analyses of AP patient datasets and experiments in mouse models demonstrated that necroptosis was significantly enriched in AP, accompanied by marked activation of the RIPK1/RIPK3/MLKL pathway. Network pharmacology analysis identified nine potential therapeutic targets of CCM associated with AP, among which RIPK1 was highlighted as a critical candidate. Furthermore, molecular docking, MD simulations, and CETSA demonstrated a strong and stable binding interaction between CCM and RIPK1. Experiments conducted in pancreatic acinar-like cells showed that CCM dose-dependently reduced the phosphorylation levels of RIPK1, RIPK3, and MLKL, with high-dose CCM markedly suppressing necroptosis. CONCLUSION: These findings indicate that CCM suppresses necroptosis, a critical pathological form of cell death in the development of AP, by modulating the RIPK1/RIPK3/MLKL pathway, thereby providing the first evidence for its therapeutic potential in AP.