Yindan Pinggan Capsules alleviates acetaminophen-induced liver injury by modulating ferroptosis via the system Xc−/GSH/GPX4 axis and the LPCAT3–ACSL4 pathway

Abstract

Drug-induced liver injury is an increasingly serious health problem, with a relatively high prevalence in China. Excessive acetaminophen (APAP) intake is a common cause of acute liver injury. Mechanistically, the toxic APAP metabolite N -acetyl- p -benzoquinone imine (NAPQI) depletes hepatic glutathione (GSH), thereby triggering oxidative stress. Yindan Pinggan Capsules (YDPG), a traditional Chinese medicine prescription, is used for clearing heat and promoting diuresis. However, their protective effects against APAP-induced liver injury (AILI) and the underlying molecular mechanisms remain unclear. This study aimed to evaluate the protective effect of YDPG against AILI and to elucidate the underlying mechanisms. Adult male mice were administered YDPG by oral gavage once daily for 7 days. One hour after the final YDPG administration, APAP at 300 mg/kg was injected intraperitoneally to establish a mouse model of AILI. Histopathological examination confirmed that YDPG significantly ameliorated APAP-induced hepatic necrosis and inflammation. To explore the molecular mechanisms underlying these effects, Gene Expression Omnibus data analysis, network pharmacology, and proteomics were employed. Western blot was utilized to detect the hepatic protein levels of solute carrier family 7 member 11, solute carrier family 3 member 2, glutathione peroxidase 4 (GPX4), ceruloplasmin, solute carrier family 39 member 14, acyl-CoA synthetase long-chain family member 4 (ACSL4), and lysophosphatidylcholine acyltransferase 3 (LPCAT3). YDPG significantly downregulated the overproduction of interleukin-6 and interleukin-1β in serum. Furthermore, YDPG effectively attenuated the APAP-induced elevation of serum alanine aminotransferase and aspartate aminotransferase levels. Comprehensive proteomics analysis revealed that YDPG specifically modulated the ferroptosis pathway. Western blot and immunohistochemistry confirmed the regulatory effects of YDPG on key proteins involved in ferroptosis. The findings of this study demonstrate that YDPG can alleviate hepatic injury and inhibit the progression of AILI in mice. These protective effects are attributed to YDPG’s ability to suppress the system Xc − /GSH/GPX4 axis and modulate the LPCAT3–ACSL4 pathway. Consequently, this research highlights the potential of YDPG as a therapeutic option for AILI.