基于回肠顶端钠依赖性胆酸转运体探究栀子水提物对小鼠胆汁淤积的治疗作用

Effect of Gardenia jasminoides water extract on cholestasis in mice based on apical sodium-dependent bile acid transporter of ileum

目的 基于回肠顶端钠依赖性胆酸转运体(apical sodium-dependent bile acid transporter,ASBT)考察栀子水提物对α-异硫氰酸萘酯(α-naphthylisothiocyanate,ANIT)诱导的胆汁淤积小鼠的治疗作用,并筛选潜在的活性化合物。方法C57BL/6小鼠随机分为对照组、栀子对照(240 mg/kg)组、ANIT组及栀子水提物低、中、高剂量(60、120、240 mg/kg)组和熊去氧胆酸(60 mg/kg)组,每组10只。给予药物进行干预,2次/d,连续14 d;于第12天单次ig ANIT(50 mg/kg)诱导小鼠胆汁淤积模型。采用栀子水提物干预ANIT诱导的小鼠小肠上皮MODE-K细胞;测定血清生化指标及肝脏、血清、胆囊、回肠及粪便中胆酸盐含量;测定小鼠回肠及MODE-K细胞ASBT及法尼酯X受体(farnesoid X receptor,FXR)的表达;采用原位肠灌流及外翻肠囊法测定回肠组织及肝门静脉中牛磺胆酸钠(taurocholate sodium,TCA)含量;采用定性代谢组学技术测定给予栀子水提物2 h后血清及回肠组织中的化合物。结果 与ANIT组比较,栀子水提物能缓解肝脏病变,减少肝脏胆酸盐及回肠TCA含量(P<0.05、0.01),增加粪便胆酸盐的含量(P<0.05);增加小鼠回肠及MODE-K细胞FXR表达(P<0.05、0.01、0.001),减少ASBT的表达(P<0.01、0.001)。同时,筛选出潜在的活性化合物37个。结论 栀子水提物可通过激活回肠FXR而抑制回肠ASBT的表达和功能,减少回肠胆酸盐重吸收并促进粪便排泄,减少肝脏胆酸盐蓄积,从而缓解ANIT诱导的小鼠肝内胆汁淤积。

Objective To investigate the effect of Gardenia jasminoides water extract on α-naphthyl isothiocyanate(ANIT)-induced cholestasis in mice based on apical sodium dependent bile acid transporter(ASBT) of ileum, and screen the potential active compounds.Methods C57BL/6 mice were randomly divided into control group, G. jasminoides control group(240 mg/kg), ANIT group, G.jasminoides water extract low-, medium-and high-dose(60, 120, 240 mg/kg) groups and ursodeoxycholic acid(60 mg/kg) group, with 10 mice in each group. Drug intervention was given twice a day for 14 d;Cholestasis model of mice was induced by single ig ANIT(50 mg/kg) on 12th day. G. jasminoides water extract was used to interfere with ANIT-induced MODE-K cells;Serum biochemical indexes and bile salt content in liver, serum, gallbladder, ileum and feces were measured;The expressions of ASBT and farnesoid X receptor(FXR) in ileum of mice and MODE-K cells were measured;The contents of sodium taurocholate(TCA) in ileum and hepatic portal vein were determined by in situ intestinal perfusion and everted intestinal sac method;Qualitative metabonomics was used to determine the compounds in serum and ileum tissue after 2 h of ig G. jasminoides water extract. Results Compared with ANIT group,G. jasminoides water extract could alleviate liver diseases, reduce the content of bile salts in liver and TCA in ileum(P < 0.05, 0.01),and increase the content of fecal bile salts(P < 0.05);G. jasminoides water extract increased the expression of FXR in ileum of mice and MODE-K cells(P < 0.05, 0.01, 0.001), and decreased the expression of ASBT(P < 0.01, 0.001). At the same time, 37 potential active compounds were screened. Conclusion G. jasminoides water extract can inhibit the expression and function of ASBT by activating FXR in ileum, reduce bile salt reabsorption in ileum, promote fecal excretion, and reduce hepatic bile salt accumulation,thereby relieving ANIT-induced intrahepatic cholestasis in mice.