Huo Po Xia Ling Decoction Exerts Preventive and Improvement Effects on Gastric Precancerous Lesions by Remodeling Gut Microbiota and Associated Metabolites

Huo Po Xia Ling Decoction (HPXLD) is a classical formulation in traditional Chinese medicine (TCM), and has demonstrated potential in managing gastric precancerous lesions (GPL). However, the mechanisms responsible for these effects remain unclear. The efficacy and associated mechanisms of HPXLD were investigated in a murine model of GPL by examining alterations in gut microbiota composition and metabolic profiles associated with disease progression and treatment. The chemical constituents of HPXLD were first characterized via ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS). A GPL mouse model was then developed using N-methyl-N-nitro-N-nitrosoguanidine (MNNG) administration in conjunction with irregular feeding protocols. Gastric tissue morphology was assessed using H&E staining and immunohistochemical techniques. The structure and diversity of intestinal microbiota were assessed by 16S rDNA sequencing, while untargeted metabolomics and MetOrigin-based pathway tracing were employed to identify treatment-associated metabolic shifts. Key differential bacterial taxa and metabolites were screened, and correlation analyses were performed to link microbiota and metabolites with inflammatory markers. HPXLD treatment significantly promoted gastric mucosal healing, attenuated glandular atrophy and inflammatory infiltration, and suppressed the expression of pro-inflammatory cytokines. 16S rDNA sequencing revealed that HPXLD substantially reshaped the intestinal microbial landscape, characterized by increased relative abundances of bacteroidota, actinobacteria, and proteobacteria, and a concurrent reduction in firmicutes. The intervention also enhanced the proliferation of probiotic taxa such as bifidobacterium, while restoring overall microbial diversity. Furthermore, metabolomic analysis combined with Metorigin analysis identified tryptophan metabolism as the pivotal therapeutic pathway. These results suggest that HPXLD exerts significant therapeutic effects in GPL by rebalancing the gut microbiota, modulating tryptophan metabolism, and reducing inflammation. These findings not only validate the clinical potential of HPXLD in managing GPL but also underscore the significant contribution of the gut microbiota-metabolite-inflammation axis in the pathogenesis and treatment of precancerous gastric conditions.