Phenylethyl Isothiocyanate Mitigates Drug-Induced Liver Injury in Mice by Inhibiting Hepatocyte Pyroptosis through the NLRP3-Caspase-1-GSDMD Pathway

Ning An ¹ XiaoRu Wang ¹ Meng Cheng ¹ Mei Bai ¹ Jia Qin ¹ jincai Cheng ² Qiang Xu ^1,3* Xuefeng Wu ^1*

• ¹ State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing, Jiangsu Province, China
• ² Drug R&D Institute, JC (Wuxi) COMPANY, Inc., Wuxi, Liaoning Province, China
• ³ Nanjing University, Nanjing, China

Drug-induced liver injury (DILI) represents a specific category of adverse liver reactions resulting from drug exposure. The pathological mechanisms underlying DILI are complex and multifaceted, and they remain incompletely understood. One potential common mechanism associated with various drug-induced instances of DILI is cell pyroptosis. In this study, we developed a mouse model of DILI using triptolide and examined the alleviating effects of the natural compound phenethyl isothiocyanate (PEITC) on DILI through a distinct mechanism involving the suppression of hepatocyte pyroptosis. Our findings revealed that PEITC administration at doses from 5 to 20 mg/kg significantly ameliorated liver tissue damage, restored normal liver architecture, decreased serum transaminase levels (AST and ALT), and normalized hepatic metabolic function in mice with DILI. The protective effects of PEITC are closely associated with its ability to directly inhibit hepatocyte pyroptosis. Specifically, PEITC effectively reversed the upregulation of the inflammasome NLRP3 in the livers from mice treated with triptolide, as well as the cleavage of Caspase-1 and Gasdermin D (GSDMD). In vitro experiments using cultured hepatocytes demonstrated that PEITC directly protects the cells by inhibiting the expression and activation of NLRP3, Caspase-1, GSDMD, and other proteins involved in the pyroptosis signaling pathway. Furthermore, electron microscopy confirmed that PEITC inhibits the formation of membrane pores in hepatocytes triggered by pyroptosis. In summary, our study identifies a novel therapeutic target for DILI and offers new insights into the clinical application of PEITC.