This experiment was conducted to determine the effects of Broussonetia papyrifera leaf water extract on the immunity and gut microflora of cyclophosphamide-induced immunosuppressed mice. Forty female ICR mice were randomly divided into five groups (n = 8): the blank control (BC) group, the cyclophosphamide-induced immunosuppression model (CTX) group, and the low (BPL), medium (BPM), and high (BPH) dose B. papyrifera extract treatment groups. Broussonetia papyrifera leaf water extract was administered intragastrically to the BPL, BPM, and BPH groups at 2, 4, and 8 g/kg body weight, respectively, once a day for 14 d. From day 12, all except the BC group mice were injected with 50 mg/kg cyclophosphamide once a day for 3 d. Administering B. papyrifera leaf extract substantially enhanced the contents of white blood cells, lymphocytes, monocytes, neutrophils, IgG, and IgM, compared with the CTX group. The B. papyrifera leaf extract also restored the gut microbiota composition by decreasing the relative abundance of Lactobacillus, Lachnospiraceae_NK4A136 group, Roseburia, Lachnospiraceae_uncultured, Lachnoclostridium, and Anaerotruncus, and increasing the relative abundance of Bacteroidales_S24-7 group norank, Desulfovibrio, Akkermansia, Enterorhabdus, Blautia, and Romboutsia in the cyclophosphamide-induced immunosuppressed mice. These findings suggest that B. papyrifera leaf extract can be used as an immunomodulator of the gut microbiota, with the potential to promote animal health.

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.