Long-term use of Sorafenib (SOR) in patients with hepatocellular carcinoma will lead to drug resistance, leading to diarrhea, weight loss and other adverse reactions. Cistanche deserticola, as a homologous substance of medicine and food, its total glycosides (TG) have anti- hepatocellular carcinoma activity. This paper aimed to explore the anti-hepatocellular carcinoma effect of TG combined with SOR on mice and its mechanism. In this study, according to the previous research of experts, the dosage of 30mg/kg SOR was selected, aiming at giving consideration to the anti-tumor effect and reducing toxicity. Specifically, HepG2 cells were used to establish a subcutaneous tumorigenic model. Histological changes, oxidative stress, immune regulation and intestinal metabolism and microbial composition of mice were detected. It was found TG did not affect the normal growth of mice without any toxic side effects and could synergize with SOR to inhibit tumor growth. HE results found mice in TG group had more complete hepatocyte structure and less pathological changes in liver. The tumor tissue cells became rare and necrotic. TUNEL staining results revealed that TG could significantly promote apoptosis of HepG2 cells, but at a lower level than SOR. In addition, the content of CAT (Catalase), SOD (Superoxide Dismutase) and ALT/GPT (Glutamic-pyruvic transaminase) increased, AST/GOT (Aspartate aminotransferase) and AKP/ALP (Alkaline phosphatase) decreased, indicating TG could effectively reduce liver lesions. TG could also decrease the content of Interleukin-2 (IL-2), Interleukin-17 (IL-17), Tumour necrosis factor-α (TNF-α), etc., and increase interferon-γ (IFN-γ), granulocyte-macrophage colony stimulating factor (GM-CSF). Meanwhile, immunohistochemistry was used to observe the decreased expression of Ki67, β-catenin and Dishevelled (Dsh) proteins and the increased GSK-3β in the tumor tissues, it was presumed that TG exerted its anti-hepatocellular carcinoma activity through the Wnt/β-catenin signaling pathway. Most importantly, TG could synergize with SOR to regulate metabolic levels in the gut, balance intestinal microbial composition, and increase the abundance of beneficial intestinal bacteria. In conclusion, the results showed that TG (64mg/kg) could assist SOR (30mg/kg) to inhibit the development of hepatocellular carcinoma, provide some theoretical basis and technical support. Cistanche deserticola, as a homologous substance of medicine and food, its combination with SOR can provide a new idea for the treatment of hepatocellular carcinoma and its future application potential is worth further exploration and development.