目的 基于中医“五脏苦欲补泻”中“肝欲散”理论，探究黄连Coptidis Rhizoma分别配伍酸味药乌梅Mume Fructus与辛味药吴茱萸Evodiae Fructus治疗胆汁淤积的效应差异及生物学机制，以阐释“酸泻”与“辛补”的科学内涵。方法 将36只雄性SD大鼠随机分为对照组、模型组、黄连组、黄连-乌梅组、黄连-吴茱萸组和熊去氧胆酸组。采用α-萘异硫氰酸酯构建胆汁淤积大鼠模型，给予药物干预后，通过血清生化指标碱性磷酸酶（alkaline phosphatase，ALP）、γ-谷氨酰转移酶（γ-glutamyl transferase，GGT）、总胆红素（total bilirubin，TBIL）、总胆汁酸（total bile acid，TBA）和组织病理学评估各给药组抗胆汁淤积效应；通过血清中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-β）、IL-6水平评估抗炎效应；通过血清中超氧化物歧化酶（superoxide dismutase，SOD）活性及丙二醛（malondialdehyde，MDA）、活性氧（reactive oxygen species，ROS）水平评估抗氧化应激效应；通过血清中总胆固醇（total cholesterol，TC）、三酰甘油（triglyceride，TG）、低密度脂蛋白胆固醇（low-density lipoprotein cholesterol，LDL-C）水平评估调血脂效应；运用LC-MS非靶向代谢组学技术分析粪便代谢物，筛选差异代谢物，并进行京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）富集分析和基因集富集分析（gene set enrichment analysis，GSEA）。结果 与模型组比较，各给药组血清生化指标水平显著降低（P＜0.05、0.01、0.001），且肝组织炎细胞浸润、肝细胞肿胀及局灶性坏死等现象明显减轻。与黄连组比较，黄连-乌梅组抗炎和抗氧化应激效应显著增强（P＜0.05、0.01），黄连-吴茱萸组调血脂效应显著增强（P＜0.05）。代谢组学分析显示，黄连配伍乌梅后抗炎效应和抗氧化应激效应增强与初级胆汁酸生物合成通路的调控有关；黄连配伍吴茱萸后调血脂效应的增强与亚油酸代谢通路的调控有关。结论 黄连在配伍乌梅或吴茱萸后对胆汁淤积大鼠均有确切的干预效应，且呈现出“酸泻”与“辛补”差异，对初级胆汁酸生物合成通路和亚油酸代谢通路的调控是导致该效应差异的潜在机制。

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.

R285.5

国家自然科学基金资助项目（82374331）；四川省科技创新苗子工程培育项目（MZGC20240077）