Multi-omics Analysis Reveals the Mechanism of Huaganjian in Alleviating Cholestatic Liver Fibrosis

Huaganjian (HGJ) is a traditional Chinese medicinal formula with liver-protective effects. However, the pharmacological mechanisms of the effects of HGJ on cholestatic liver fibrosis (CLF) are yet to be clarified. To evaluate the effects of HGJ on CLF and elucidate the underlying mechanisms, C57BL/6J mice were fed a 0.1% 3, 5-diethoxycarbonyl-1, 4-dihydrocollidine (DDC) diet to induce CLF. The efficacy of HGJ was evaluated by measuring the biochemical indicators of liver function, fibrosis, and histology. The underlying mechanisms were investigated using an integrated multi-omics approach, including fecal 16S rRNA sequencing, serum metabolomics, and hepatic transcriptomic analysis. The findings were further validated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting (WB). HGJ significantly alleviated liver injury, cholestasis, and fibrosis. Microbiome analysis revealed that Bifidobacterium, Turicibacter, and Clostridium_sensu_stricto_1 abundances were positively correlated with liver injury and fibrosis marker levels, and these abundances decreased following HGJ treatment. Metabolomic analysis identified 531 differential metabolites, including 299 upregulated and 232 downregulated metabolites, following HGJ intervention. Hepatic transcriptomic analysis revealed 164 differentially expressed genes, including 102 upregulated and 62 downregulated genes. Integrated multi-omics analysis revealed that HGJ alleviated CLF by modulating the glycine/serine/threonine metabolism pathway. RT-qPCR and western blotting experiments confirmed that in this pathway, aminolevulinic acid synthase 1 levels decreased, whereas serine dehydratase and serine dehydratase-like levels increased after HGJ treatment. Overall, HGJ effectively alleviated CLF, and its mechanisms of action were closely linked to the regulation of the glycine/serine/threonine metabolism pathway.