基于胆汁酸代谢组学探讨芍药苷改善α-萘异硫氰酸酯诱导胆汁淤积的作用机制

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（dire...

Mechanism of paeonilforin on ameliorating α-naphthylisothiocyanate-induced cholestasis based on bile acid metabolomics

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.