Physcion-8-O-β-D-monoglucoside protects hepatocytes from TNF-α-mediated apoptosis by suppressing the PI3K/AKT /NF-κB signaling pathway

Abstract Physcion-8-O-β-D-monoglucoside (PMG) is one of the active ingredients of Radix et Rhizoma Rhei, which had been used for treating liver disorders for hundreds of years in China. However, the hepatoprotective effects of PMG remain poorly understood. This study aimed to investigate the mechanism of the protection effects of PMG on Tumor necrosis factor-α (TNF-α)-induced hepatotoxicity. We developed both in vitro and in vivo models of liver injury to assess the protective effects of PMG against TNF-α-induced hepatotoxicity. The in vitro model employed TNF-α/actinomycin D in AML-12 cells, while the in vivo model utilized intraperitoneal injection of carbon tetrachloride (CCl₄) in mice. Interactions of PMG and TNFR1 (the receptor of TNF-α) were explored by molecular docking. AAV resuspension was administered before PMG treatment via intravenous injection to overexpress TNF-α in the CCl₄-induced mice. Liver injury markers were examined, and the associated changes were detected using CCK8, Hoechst staining, Western blotting, and other molecular assays. PMG effectively reversed the morphological changes, restoring cell shape and structure in TNF-α injured cells. PMG protected against hepatotoxicity in vitro and in vivo. PMG and TNFR1 maintained robust binding activities. TNF-α overexpression counteracted the hepatoprotective effects of PMG, attenuating its influence on IL-6, AST, ALT, apoptosis, and the inactivation of the PI3K/AKT/NF-κB signaling pathway. PMG protected against TNF-α-induced hepatotoxicity by regulating the PI3K/AKT/NF-κB signaling pathway through TNF-α inhibition, suggesting that PMG holds potential as a novel therapeutic agent for acute liver injury.