Background <p>Severe acute pancreatitis (SAP) often induces colitis, but the mechanisms remain unclear. Calcium dyshomeostasis is implicated in this process. This study investigates SARAF, a calcium influx channel inhibitor, in SAP-induced colitis.</p> Methods <p>The role of SARAF in SAP-induced colitis was investigated using LPS-stimulated NCM460 colonic cells and a caerulein-induced rat model. Techniques included Western blot, RT-qPCR, ELISA, calcium imaging, and histopathological analysis. SARAF was overexpressed to evaluate its effects on cell viability, inflammatory responses, and the H₂S/CSE/CBS and NO/iNOS pathways.</p> Results <p>SARAF expression was significantly downregulated in inflammatory conditions. SARAF overexpression reduced intracellular Ca²⁺ overload, decreased pro-inflammatory cytokines, and enhanced cell viability. It suppressed the H₂S/CSE/CBS and NO/iNOS pathways at transcriptional and protein levels. In vivo, SARAF administration alleviated colonic histopathological damage, improved motility, reduced bowel movement frequency, and decreased serum NO levels. Histological scores showed significant improvement in inflammatory cell infiltration, crypt destruction, and epithelial damage.</p> Conclusion <p>SARAF protects against SAP-induced colitis by modulating the H2S/CSE/CBS and NO/iNOS pathways and restoring calcium homeostasis, suggesting its potential as a therapeutic target for colitis intervention.</p>