Multi-omics Profiling Reveals the Effects of Xihuangcao on Hepatic Fibrosis Attenuation through Gut Microbiota and Metabolic Reprogramming

In various chronic liver diseases, hepatic fibrosis represents a frequent pathological event. This process involves the activation of hepatic stellate cells, along with the consequent excessive deposition of extracellular matrix. Emerging evidence highlights the contribution of gut microbiota dysbiosis and intestinal barrier dysfunction to its progression, underscoring the potential of targeting the liver-gut axis. Xihuangcao, a traditional Chinese medicinal herb used for acute cholecystitis and jaundice hepatitis, exhibits antifibrotic activity, but its underlying mechanisms remain unclear. In this study, hepatic fibrosis models were established to evaluate the antifibrotic effects of 3 botanical sources of Xihuangcao (Isodon lophanthoides (Buch.-Ham. ex D.Don) H.Hara, Isodon lophanthoides var. graciliflorus (Benth.) H.Hara, and Isodon serra Kudô) through integrating histopathological, transcriptomics, metabolomics, 16S ribosomal RNA (rRNA) sequencing, and network pharmacology analyses. The results indicated that all 3 botanical sources of Xihuangcao markedly alleviated hepatic fibrosis and reduced collagen deposition. Transcriptomic analysis revealed that Xihuangcao down-regulated profibrotic mediators (Toll-like receptor 5, transforming growth factor-β, and platelet-derived growth factor) and modulated mitogen-activated protein kinase, steroid biosynthesis, bile secretion, and retinol metabolism pathways, highlighting its coordinated suppression of inflammation-driven fibrogenic activation in hepatic fibrosis. Liver metabolomics revealed Xihuangcao's effect on restoration of energy metabolism, amino acid turnover, and redox homeostasis. Xihuangcao also corrected fibrosis-induced gut dysbiosis, characterized by reduced Verrucomicrobia and increased S24-7 abundance, while suppressing pathogenic genera such as Clostridium and Prevotella. Moreover, Xihuangcao decreased profibrotic metabolites (androstan-3α,17β-diol) and suppressed choline-related metabolic pathways associated with fibrogenic signaling. Network pharmacology and experimental validation further identified PI3K/Akt inhibition as a key mechanism suppressing hepatic stellate cell activation. In summary, this study, based on integrative multi-omics analyses (metabolomics, transcriptomics, and 16S rRNA sequencing), found that Xihuangcao alleviates liver fibrosis by modulating inflammatory responses, metabolic disorders, and gut microbiota dysbiosis via the gut-liver axis. Furthermore, network pharmacology combined with in vitro experiments revealed that inhibition of hepatic stellate cell activation through the PI3K/Akt pathway may be the critical mechanism underlying its therapeutic efficacy.