Targeting ACOT1-SLC25A5 Axis attenuates LPS-induced acute liver injury by inhibiting ferroptosis and inflammatory responses.

Abstract

Sepsis associated acute liver injury is a major contributor to multiple organ dysfunction and mortality in critically ill patients. ACOT1 has emerged as a promising therapeutic target, demonstrated to inhibit disseminated intravascular coagulation through its anti-ferroptosis activity. However, its specific function and mechanistic role in the context of during endotoxemia-induced liver injury have not been elucidated. This study demonstrates that ferroptosis was markedly activated in both lipopolysaccharide (LPS)-stimulated AML12 hepatocytes and LPS-induced acute liver injury mouse models. RNA sequencing and bioinformatics analyses identified ACOT1 as the most significantly downregulated genes following LPS stimulation. Functionally, ACOT1 overexpression upregulated GPX4 and SLC7A11 expression, reduced mitochondrial ROS and Fe2+ accumulation, which ultimately attenuated LPS-induced hepatocyte ferroptosis. Mechanistically, ACOT1 overexpression activated PPARγ, thus suppressing NF-κB pathway activation and inflammatory responses thereafter. IP-MS and immunofluorescence staining confirmed that SLC25A5 interacts with ACOT1 and functions as a molecular partner in regulating hepatocyte ferroptosis by reducing mitochondrial ROS and Fe2+ levels. Protein interaction analysis between ACOT1 and SLC25A5 showed a Rosetta score of -271.16 kcal/mol, indicating strong binding affinity. Collectively, our results identified ACOT1 as a novel inhibitor of ferroptosis in LPS-induced acute liver injury. ACOT1 exerts protective effects by modulating the PPARγ/NF-κB signaling axis and cooperating with SLC25A5 to regulate oxidative stress-driven ferroptosis. These results highlight ACOT1 as a potential regulator for inflammatory liver injury.