<p>Acute pancreatitis (AP) is a rapid-onset inflammatory disorder of the pancreas, characterized by premature activation of pancreatic digestive enzymes and the development of systemic inflammatory responses. Severe AP is associated with a mortality rate of 10–40%, highlighting the urgent need for effective and targeted therapeutic interventions. Obtusifolin (OBT), a natural compound from <i>Senna obtusifolia</i>, exhibits anti-inflammatory and wound-healing properties. The current study aimed to investigate the effect of OBT against inflammation and tissue injury associated with AP. In vitro, lipopolysaccharide (LPS) and TGF-β-induced differentiation models were employed to investigate the anti-inflammatory and anti-fibrotic effects of OBT in pancreatic stellate cells (PSCs) and PANC-1 cells. In vivo, a cerulein-induced acute pancreatitis mouse model was employed to evaluate the therapeutic potential of OBT through histopathological analysis, ELISA, immunohistochemistry, and western blotting. In vitro results revealed that OBT treatment significantly suppressed the LPS/TGF-β-induced pro-inflammatory and ECM marker expression in PSCs and PANC-1 cells, respectively. In mice, cerulein induction notably increased the pancreatic edema, acinar necrosis, inflammatory infiltration, hemorrhage in tissues of cerulein control, on the other hand, treatment with OBT significantly attenuated the same. Further, OBT treatment significantly reduced the cerulein-induced elevation of inflammatory marker expression (<i>Tnfa, Ccl2, Cxcl10,</i> and <i>Il6</i>), serum α-amylase, β-amylase, and IL-1β levels in a dose-dependent manner. Furthermore, immunohistochemistry and western blot analysis confirmed that OBT ameliorates pancreatitis by modulating the NFκB signaling. These results indicate that obtusifolin attenuates acute pancreatitis by inhibiting inflammatory responses and preserving pancreatic tissue integrity, supporting its potential as a therapeutic candidate for managing acute pancreatitis.</p> Graphical Abstract <p></p>