This study aimed to investigate the hepatoprotective effects of total flavones from Hippophae rhamnoides L.(TFH) against carbon tetrachloride CCl₄-induced acute liver injury (ALI) in mice. ALI was induced in mice via intraperitoneal injection of 0.2% CCl₄-olive oil solution. Hepatic function, histopathological morphology, and biochemical markers related to oxidative stress, and inflammatory cytokines were evaluated. The mRNA expression levels of key genes were quantified using RT-qPCR. Compared with the control group, the CCl₄-induced model group exhibited significantly elevated serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and malondialdehyde (MDA), along with increased hepatic levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) (P < 0.01). Conversely, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in liver tissues were markedly reduced (P < 0.01). Histopathological analysis revealed extensive hepatocyte necrosis, inflammatory infiltration, hepatic enlargement, and a significantly higher liver index (P < 0.01). Furthermore, the mRNA expression levels of TNF-α, IL-6, and signal transducer and activator of transcription 3 (STAT3) were significantly upregulated in the model group (P < 0.01). In contrast, TFH treatment dose-dependently attenuated these alterations. Specifically, TFH administration reduced serum AST, ALT, and MDA levels, decreased hepatic TNF-α and IL-6 concentrations, and enhanced SOD and GSH-Px activities (P < 0.01). Histopathological improvements, including reduced hepatocyte necrosis and inflammatory infiltration, were observed. RT-qPCR analysis confirmed the downregulation of TNF-α, IL-6, and STAT3 mRNA expression in TFH-treated groups. TFH exerts hepatoprotective effects against CCl₄-induced ALI by modulating inflammatory cytokine expression, suppressing the activation of the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway, and downregulating the expression of downstream gene such as STAT3.