Disulfiram alleviates immune-mediated liver injury by inhibiting pyroptosis in hepatocytes through the NF-κB pathway

Abstract

Immune-mediated liver injury (ILI) is a condition characterized by inflammation and cell death in the liver. Disulfiram, an FDA-approved drug for alcohol aversion, shows potential as a therapeutic agent in liver diseases, although its effects on immune liver injury remain unclear. This study aims to investigate the therapeutic effects of disulfiram using a mouse model of ILI induced by concanavalin A (ConA) and AML12 hepatocytes in vitro. Various techniques were employed, including Western blotting, qRT-PCR, cell viability assays, histopathological evaluations, immunohistochemistry, TUNEL staining, caspase-1 activity assays, cytokine detection, Cellular Thermal Shift Assay, and EdU proliferation assays. The results demonstrate that the ConA-induced ILI model exhibits significant liver damage and cellular pyroptosis, with disulfiram administered at specific concentrations markedly reducing the inflammatory response. Moreover, disulfiram attenuates pyroptosis in ConA-induced ILI and reduces cell injury in AML12 hepatocytes triggered by ConA and LPS + ATP, while facilitating post-injury cell proliferation. Mechanistically, the anti-pyroptosis effects of disulfiram are associated with the inhibition of the NF-κB signaling pathway in vitro. These results suggest that hepatocellular pyroptosis plays a pivotal role in the pathogenesis of ILI, and that disulfiram alleviates ILI symptoms by modulating the NLRP3/Caspase-1/GSDMD-mediated classical pyroptosis pathway through the NF-κB signaling cascade. Subsequent investigations will explore the impact of disulfiram in diverse liver injury models and its synergistic effects with other drugs to improve therapeutic outcomes. Additionally, clinical trials are imperative to validate these findings in humans and establish disulfiram as a standard treatment for ILI, thereby paving the way for innovative therapeutic approaches.