Jiedu Huayu Extract Alleviate Acute Liver Failure via Promotion of GPX4 Expression and Inhibition of D-GalN/LPS-Induced Ferroptosis

Objective This study aims to explore the potential mechanisms of Jiedu Huayu granules (JDHY) mitigate D-galactosamine (D-GalN) and lipopolysaccharide (LPS)-induced acute liver failure (ALF) in a cell damage model. Methods ALF was modeled using various concentrations of D-GalN + LPS. JDHY-medicated serum at different concentrations was then co-cultured with the cell model in proportion. The best concentration and time of JDHY-medicated serum intervention were determined by Cell Counting Kit-8, Alanine Aminotransferase (ALT), and Aspartate Aminotransferase (AST). Western blot was used to assess the expression of Ferritin Heavy Chain 1 (FTH1), Transferrin Receptor 1(TfR1), Glutathione Peroxidase 4 (GPX4), Lysyl Oxidase (LOX), Prostaglandin-Endoperoxide Synthase 2(PTGS2). Malondialdehyde was analyzed for cell lipid peroxidation, and enzyme-linked immunosorbent assay was used to detect glutathione, Tumor Necrosis Factor-alpha, Interleukin-10, Interleukin-6 expression, and liver function indicators (ALT, AST). Additionally, GPX4 was knocked down using cell transfection, and the molecular mechanisms of JDHY in treating ALF were explored through Western blot, PCR, and enzyme-linked immunosorbent assay. Results The appropriate dose and time of D-GalN/LPS-induced ALF (10 mg/mL D-GalN + 1 μg/mL LPS for 48 h) and the optimal intervention concentration of JDHY-medicated serum (15%) were determined through ALT, AST, and Cell Counting Kit-8 assays. JDHY treatment reduced ALT and AST levels, alleviated cell lipid peroxidation, and inhibited ferroptosis. The mechanism involves JDHY enhancing the antioxidant capacity in liver cells by increasing the expression of GPX4 and glutathione, regulating ferroptosis proteins (downregulating TfR1, upregulating FTH1), inhibiting LOX and PTGS2, and suppressing inflammation (downregulating Tumor Necrosis Factor-alpha and Interleukin-6, upregulating Interleukin-10). In addition, GPX4 knockdown experiments revealed that knocking down GPX4 worsened ALF, while JDHY can alleviate ALF by promoting GPX4 expression and enhancing the antioxidant capacity of liver cells. Conclusion JDHY enhance GPX4 expression and reduce lipid peroxidation in liver cells affected by ALF, protecting liver cell, alleviating inflammatory, and inhibiting ferroptosis.