Abstract Aims Based on the gut microbiota and plasma metabolites, the underlying mechanism was analysed for the anti‐hepatocellular carcinoma (HCC) effects of Ganfule capsule (GFL) in the study. Methods and results The UPLC‐Q‐TOF/MS results showed that 13 key compounds were identified in GFL and the major active ingredients included amygdalin, saikosaponin A, astragaloside I, etc. The nude mice received HepG2 injection, and GFL showed lower volume and weight of the tumour. In addition, the apoptosis proteins (Bax and Bcl2) were altered in response to GFL treatment, and apoptosis cells were increased, indicating an anti‐HCC effect. Interestingly, 16S rDNA results showed that GFL treatment improved gut microbiota diversity and compositions, especially for the beneficial bacteria, such as Bacilli, Lactobacillales, Lactobacillus, Lactobacillaceae, Firmicutes, Lactobacillus_reuteri and Lactobacillus_gasseri. Metabonomics further identified 426 metabolites and 343 metabolites variation in the positive and negative ion modes after GFL treatment, which might be associated with amino acid, lipid metabolism and carbohydrate metabolism pathways, indicating these metabolites might involve in the protective role of GFL in HCC. Correlation analysis showed a significant relationship between gut microbiota and plasma metabolites. Conclusion In conclusion, GFL exerted an anti‐HCC effect in the nude murine model, which might be associated with microbial and metabolic improvements. Significance and impact of the study This study is the first to report the anti‐HCC effect of GFL associated with gut microbiota and plasma metabolites. GFL may improve the gut microbiota structure, such as increasing probiotics – Lactobacillus. It also provides a new strategy for the scientific demonstration of the modernization of traditional Chinese medicine.
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