Ferroptosis and Lipid Peroxidation Participate in Valproic Acid‐Induced Hepatotoxicity via the Long‐chain Acyl‐CoA Synthetase 4/Glutathione Peroxidase 4 Pathway

Valproic acid (VPA) is a commonly prescribed antiepileptic drug, with hepatotoxicity being one of its most frequent and severe adverse effects. The underlying mechanisms of VPA-induced hepatotoxicity remain elusive. Thus, this study aimed to investigate the involvement of ferroptosis in VPA-induced hepatotoxicity in vivo and in vitro. C57BL/6 J mice and HepG2 cells were treated with VPA to establish VPA-induced hepatotoxic models. The results demonstrated that VPA not only induced hepatic steatosis but also elevated liver biochemical and oxidative stress indicators, suggesting that VPA-induced hepatotoxicity affects hepatic iron metabolism. Moreover, VPA treatment altered the expression of ferroptosis-related proteins and lipid peroxides, indicating that ferroptosis contributed to VPA-induced hepatotoxicity. To investigate the role of ACSL4, a pivotal enzyme in lipid peroxidation during ferroptosis, in VPA-induced hepatotoxicity, a study was conducted utilizing rosiglitazone (RSG), a specific inhibitor of ACSL4, to interfere with the overexpression of ACSL4 in a model mouse system. Furthermore, an in vitro approach was employed, where ACSL4 siRNA was utilized to knock down ACSL4 expression in a cellular model of VPA-induced hepatotoxicity. This dual-pronged strategy aimed at elucidating the mechanistic contributions of ACSL4 in mediating the deleterious effects of VPA on the liver. In summary, ferroptosis emerges as a novel mechanism underlying VPA-induced hepatotoxicity, and ACSL4 may serve as a crucial target in the process of VPA-induced liver injury. This study has the potential to lay the groundwork for the development of novel therapeutic strategies for treating VPA-induced liver damage.