Gentiana sino-ornata Balf. f. ethyl acetate extract alleviates high-fat diet- and tetracycline-induced liver injury in mice: insights from serum pharmacochemistry, network pharmacology, and experimental validation
Abstract
Nonalcoholic fatty liver disease (NAFLD) is one of the most prevalent chronic liver diseases worldwide, and the associated metabolic dysfunction may increase susceptibility to drug-induced liver injury (DILI). This study aimed to evaluate the hepatoprotective effects of the Gentiana sino-ornata Balf. f. ethyl acetate extract (GSEA) in mice exposed to a high-fat diet (HFD) and tetracycline. The antioxidant and anti-inflammatory activities of GSEA were evaluated in vitro and in vivo . Subsequently, the blood-absorbed ingredients of GSEA were characterized using UHPLC-HRMS, followed by network pharmacology analysis and molecular docking to elucidate the potential mechanisms underlying its hepatoprotective effects. Finally, the predicted mechanisms were experimentally validated in a mouse model of HFD- and tetracycline-induced liver injury. GSEA exhibited significant antioxidant and anti-inflammatory activities. Twenty-five blood-absorbed ingredients were tentatively identified as potentially bioactive, and the PI3K/Akt/FoxO1 signaling pathway was predicted as a pathway of GSEA against liver injury. Molecular docking further demonstrated favorable binding interactions between AKT1 and several major constituents, including sweroside, loganic acid, swertiamarin, and gentiopicroside. In vivo experiments showed that GSEA markedly alleviated HFD- and tetracycline-induced liver injury by suppressing the expression of inflammation- and glucolipid-metabolism-related genes, attenuating hepatic inflammation, modulating the gut microbiota, and partially regulating the PI3K/Akt/FoxO1 signaling pathway. Collectively, these findings provide mechanistic evidence supporting the potential application of GSEA as a natural therapeutic agent for the prevention or treatment of liver injury associated with metabolic dysfunction and tetracycline exposure.