Sepsis, defined as a systemic inflammatory response, is one of the conditions with the highest mortality rates. Crosin (CRO) is one of the active ingredients in saffron and known for its various pharmacological effects. It has been reported to have protective and healing effects on liver tissue. In this study, we aimed to investigate the protective role of CRO in lipopolysaccharide (LPS)-induced acute liver injury. 40 male Wistar Albino rats aged 8-12 weeks were randomly divided into 4 groups: Control, CRO (50 mg/kg, intraperitoneal administration for 9 days), LPS (30 mg/kg, single dose), and LPS + CRO (50 mg/kg CRO for 9 days along with a single dose of 30 mg/kg LPS). Following the experimental procedure, liver and blood samples were collected for further analyses. Histopathological analysis revealed a marked increase in liver damage in the LPS group, as evidenced by significant histopathological changes. In contrast, the liver histology in the LPS + CRO group closely resembled that of the Control and CRO groups, exhibiting substantially less damage compared to the LPS group. Immunohistochemical examinations showed a significant increase in the expressions of TLR4, HMGB1, NF-κB, TNF- α, HSP70, and HSP90 in the LPS group. However, in the LPS + CRO group, the levels of these markers were significantly lower compared to the LPS group. ELISA analyses showed a significant increase in MDA, IL-6, and TGF-β and a decrease in SOD, CAT, GSH levels in the LPS group. Conversely, in the LPS + CRO group, CRO applications exhibited a significant protective effect on these alterations. Additionally, AST, ALT, and LDH levels were significantly elevated in the LPS group, while albumin levels were lower in the LPS group. CRO applications in the LPS + CRO group were observed to have a protective effect on these parameters. We believe that CRO holds significant potential in the treatment of acute inflammatory diseases such as septic acute liver injury and should not be overlooked.