Gardenia–Astragalus–Crataegus Extract Alleviates Carbon Tetrachloride-Induced Liver Injury in Mice Through Antioxidant and Apoptosis Pathways

The incidence of liver injury has been increasing year by year, potentially progressing to liver fibrosis, cirrhosis, and even hepatocellular carcinoma, posing a threat to human health. Gardenia, Astragalus, and Hawthorn are commonly used natural products with dual purposes as both medicine and food. While extensive research has been conducted on their individual pharmacological activities, systematic studies on the synergistic effects and efficacy evaluation of their combined use in liver protection are still lacking. This study aimed to investigate the protective effects and underlying mechanisms of the compound Gardenia and Scutellaria extract (GACE) on CCl4-induced liver injury in mice. The composition and content of GACE were analyzed by HPLC. Serum biochemical, inflammatory, and hepatic oxidative stress indicators were detected using assay kits. Liver pathological changes were examined by HE staining, while apoptosis was observed via TUNEL staining. Transcriptomic and metabolomic sequencing were employed to analyze the potential mechanisms of CCl4 in alleviating liver injury. Finally, Western blotting was performed to validate the analytical results. The results demonstrated that CCl4 significantly upregulated the levels of pro-inflammatory cytokines IL-6, IL-12 and IL-2 (p < 0.05), downregulated the level of anti-inflammatory cytokine IL-10 (p < 0.05), and caused markedly elevated serum ALT and AST levels (p < 0.01). The hepatic lobule structure was nearly obliterated in liver tissue, with disordered arrangement of hepatic cords and numerous vacuoles appearing in hepatocytes. The TUNEL staining positive cell rate increased significantly compared with the blank group (p < 0.001). Meanwhile, CCl4 also significantly inhibited SOD and GSH-Px activities in liver tissue (p < 0.05), while markedly increasing MDA levels (p < 0.05). GACE significantly alleviated liver tissue damage and reduced hepatocyte apoptosis (p < 0.05). It markedly enhanced SOD and GSH-Px activities in liver tissue while decreasing ALT, AST, MDA, IL-6, IL-12, and IL-2 levels (p < 0.05), thereby mitigating hepatic oxidative damage and inflammatory responses. Integrated transcriptomic and metabolomic analyses revealed that the PI3K-Akt and MAPK signaling pathways play crucial roles in mediating GACE’s therapeutic effects against liver injury. Further validation demonstrated that GACE can attenuate CCl4-induced liver injury by inhibiting the PI3K-Akt and MAPK pathways.