目的 基于胆汁酸代谢组学及分子生物学探讨芍药苷调控胆汁酸改善胆汁淤积的作用及机制。方法 雄性SD大鼠连续7 d ig熊去氧胆酸（ursodesoxycholic acid，UDCA）或芍药苷，于实验第4天ig α-萘异硫氰酸酯（α-naphthylisothiocyanate，ANIT）建立胆汁淤积模型。通过血清生化指标、肝脏病理组织染色、胆汁酸体积流量明确芍药苷缓解胆汁淤积的药效学基础。收集大鼠胆汁进行胆汁酸代谢组学分析，收集肝脏检测法尼醇X受体（farnesoid X receptor，FXR）和胆酸盐外排泵（bile salt export pump，BSEP）表达。结果 药效学实验显示，与对照组比较，模型组大鼠血清中丙氨酸氨基转移酶（alanine aminotransferase，ALT）、天冬氨酸氨基转移酶（aspartate aminotransferase，AST）、γ-谷氨酰转移酶（γ-glutamyltranspeptidase，γ-GT）、总胆汁酸（total bile acid，TBA）、总胆红素（total bilirubin，TBIL）和直接胆红素（direct bilirubin，DBIL）水平均显著升高（P＜0.01）；给予芍药苷后，上述指标显著降低（P＜0.01），且呈剂量相关性。同时芍药苷能够改善大鼠肝组织水肿和炎性浸润，减少汇管区胆管增生，抑制肝细胞脂肪变性。胆汁酸代谢组学结果显示，与对照组比较，模型组牛磺胆酸（taurocholate acid，TCA）、牛磺α鼠胆酸（tauro-α-muricholic acid sodium，TαMCA）、胆酸（cholic acid，CA）、牛磺熊脱氧胆酸（tauroursodeoxycholic acid，TUDCA）、甘氨熊脱氧胆酸（glycoursodeoxycholic acid，GUDCA）、甘氨鹅脱氧胆酸（glycochenodeoxycholic acid，GCDCA）、甘氨胆酸（glycocholic acid，GCA）、熊脱氧胆酸（ursodeoxycholic acid，UDCA-7S）、牛磺石胆酸（taurocholic acid，TLCA）、牛磺猪脱氧胆酸（taurohyodeoxycholic acid，THDCA）、甘氨猪胆酸（glycohyocholic acid，GHCA）、甘氨脱氧胆酸（glycodeoxycholic acid，GDCA）、甘氨猪脱氧胆酸（glycohyodeoxycholic acid，GHDCA）、甘氨石胆酸（glycolithocholic acid，GLCA）、熊胆酸（ursocholic acid，UCA）、牛磺脱氧胆酸（taurodeoxycholate acid，TDCA）含量明显降低（P＜0.05、0.01），牛磺β鼠胆酸（tauro-β-muricholic acid sodium，TβMCA）含量显著升高（P＜0.01）；上述胆汁酸含量在芍药苷干预后明显回调（P＜0.05、0.01）。分子生物学验证结果显示模型组大鼠肝组织FXR和BSEP的mRNA和蛋白表达水平均明显降低（P＜0.05、0.01），经芍药苷治疗后二者表达明显上调（P＜0.05、0.01）。结论 芍药苷能够调节胆汁酸池组成，降低毒性胆汁酸含量，上调FXR和BSEP活性，恢复胆汁酸池稳态，改善ANIT诱导的胆汁淤积。

Objective To investigate the effect and mechanism of paeoniflorin on regulating bile acids to ameliorate cholestasis by metabolomics and molecular biology methods. Methods Male SD rats were consecutively ig ursodesoxycholic acid (UDCA) or paeoniflorin for 7 d, and rats was ig α-naphthylisothiocyanate (ANIT) on 4th day to establish a cholestatic model. The pharmacodynamics of paeoniflorin in relieving cholestasis was clarified by serum biochemical indices, hepatic pathological staining and bile acid flow rate. Rat bile was collected for metabolomics analysis, and hepatic samples were used to detect the expressions of farnesoid X receptor (FXR) and bile salt export pump (BSEP). Results Pharmacological results showed that compared with control group, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyltranspeptidase (γ-GT), total bile acid (TBA), total bilirubin (TBIL) and direct bilirubin (DBIL) were significantly increased in model group (P < 0.01), whereas paeoniflorin could overtly reduce the levels of the above indices (P < 0.01), and the effect was dose-dependent to a certain extent. Meanwhile, paeoniflorin ameliorated hepatic tissue edema and inflammatory infiltration, reduced bile duct hyperplasia in the portal area and inhibited hepatocellular steatosis. The results of bile acid metabolomics showed that compared with control group, the contents of taurocholate acid (TCA), tauro-α-muricholic acid sodium (TαMCA), cholic acid (CA), tauroursodeoxycholic acid (TUDCA), glycoursodeoxycholic acid (GUDCA), glycochenodeoxycholic acid (GCDCA), glycocholic acid (GCA), ursodeoxycholic acid (UDCA-7S), taurocholic acid (TLCA), taurohyodeoxycholic acid (THDCA), glycohyocholic acid (GHCA), glycodeoxycholic acid (GDCA), glycohyodeoxycholic acid (GHDCA), glycolithocholic acid (GLCA), ursocholic acid (UCA) and taurodeoxycholate acid (TDCA) were obviously reduced after ANIT induction (P < 0.05, 0.01), while the content of TβMCA was distinctly elevated (P < 0.01), while the above bile acid contents were overturned after paeoniflorin intervention (P < 0.05, 0.01). The results of molecular validation revealed that the mRNA and protein expressions of FXR and BSEP were suppressed in model group (P < 0.05, 0.01), and the expressions of both was upregulated after treatment with paeoniflorin (P < 0.05, 0.01). Conclusion Paeoniflorin can modulate the composition of bile acid pool, reduce the levels of toxic bile acid, upregulate the activities of FXR and BSEP, restore bile acid pool homeostasis, and ameliorate ANIT-induced cholestasis.

国家自然科学基金资助项目（82274187）；四川省自然科学基金资助项目（2023NSFSC0687）；中国医药教育协会2022重大科学攻关问题和医药技术难题科研课题（2022KTZ016）；中华中医药学会（2023—2025年度）青年人才托举工程项目[CACM-(2023-QNRC2-A01)]