Objective To explore the effect differences and biological mechanisms of Huanglian (Coptidis Rhizoma) combined with sour herb Wumei (Mume Fructus) and pungent herb Wuzhuyu (Evodiae Fructus) in treating cholestasis based on “liver desires dispersion” theory from “five zang-organs preferences and aversions in tonification and purgation” in traditional Chinese medicine, thereby enriching the scientific connotation of “sour purgation” and “pungent tonification”. Methods A total of 36 male SD rats were randomly divided into control group, model group, Coptidis Rhizoma group, Coptidis Rhizoma-Mume Fructus group, Coptidis Rhizoma-Evodiae Fructus group and ursodeoxycholic acid group. A cholestasis model was induced using α-naphthylisothiocyanate (ANIT). After drug intervention, the anti-cholestatic effects were evaluated by measuring serum biochemical markers sun as alkaline phosphatase (ALP), γ-glutamyl transferase (GGT), total bilirubin (TBIL), total bile acid (TBA) and histopathological examination. The anti-inflammatory effects were assessed by measuring levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 in serum. The anti-oxidative stress was evaluated by measuring activity of superoxide dismutase (SOD) and levels of malondialdehyde (MDA), reactive oxygen species (ROS) in serum. The lipid-regulating effects were evaluated by measuring levels of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in serum. LC-MS-based untargeted metabolomics was performed on fecal samples to analyze metabolites, identify differential metabolites, and conducting Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis and gene set enrichment analysis (GSEA). Results Compared with model group, the serum biochemical index levels of each treatment group were significantly reduced (P < 0.05, 0.01, 0.001), the phenomena of liver tissues inflammatory cell infiltration, liver cell swelling and focal necrosis were significantly alleviated. Compared with Coptidis Rhizoma group, Coptidis Rhizoma-Mume Fructus group demonstrated significantly enhanced anti-inflammatory and anti-oxidative stress effects (P < 0.05, 0.01), while Coptidis Rhizoma-Evodiae Fructus group showed significantly enhanced lipid-regulating effects (P < 0.05). Metabolomics analysis indicated that the enhanced anti-inflammatory and anti-oxidative stress effects following the combination of Coptidis Rhizoma with Mume Fructus were associated with the modulation of primary bile acid biosynthesis pathway, the enhanced lipid-regulating effect following the combination of Coptidis Rhizoma with Evodiae Fructus was linked to the regulation of linoleic acid metabolism pathway. Conclusion Coptidis Rhizoma, whether combined with Mume Fructus or Evodiae Fructus, exerts definite interventional effects on cholestatic rats, demonstrating the differential characteristics of “purgation with sour herbs” and “tonification with pungent herbs”. The modulation of primary bile acid biosynthesis pathway and linoleic acid metabolism pathway represents the potential mechanism underlying these differential effects.

Coptidis Rhizoma  /  Mume Fructus  /  Evodiae Fructus  /  sour-bitter heat-purging  /  pungent-opening bitter-descending  /  cholestasis  /  berberine  /  epiberberine  /  coptisine  /  palmatine  /  citric acid  /  evodiamine  /  rutaecarpin  /  limonin  /  primary bile acid biosynthesis pathway  /  linoleic acid metabolism pathway