茵甘合剂对ANIT诱导肝内胆汁淤积模型小鼠肝损伤防治作用及机制研究

摘要

目的 探讨茵甘合剂对α-萘异硫氰酸酯(ANIT)诱导肝内胆汁淤积模型小鼠肝损伤的作用.方法 正常组、模型组、茵陈组、甘草组、茵甘合剂组,每组10只小鼠.正常组小鼠灌胃0.9%氯化钠溶液10 d,模型组、茵陈组、甘草组、茵甘合剂组小鼠分别灌胃0.9%氯化钠溶液(0.2 mL/20 g)、茵陈水煎液(1.13 g/kg)、甘草水煎液(1.13 g/kg)、茵甘合煎液(茵陈1.13 g/kg、甘草1.13 g/kg)10 d,在第7天给药后1 h灌胃ANIT(100 mg/kg)造模.第10天取血样,检测血清TBil、ALT、AST、ALP和TBA;肝病理检查;RT-PCR检测Car和Pxr及其下游靶基因转录情况.结果 与模型组相比,茵甘合剂组小鼠肝脏损伤改善明显,血清TBil、ALT、AST、ALP和TBA水平分别降低22%、23%、25%、20%和30%;Car和Pxr及其下游靶基因CYP3a11、Ugt1a1、Sult2a1、Mdr2、Mrp2、Mrp3和Mrp4的mRNA的转录增加(P<0.01).结论 茵甘合剂对ANIT诱导的肝内胆汁淤积模型小鼠肝损伤具有防治作用.%Objective To investigate the protective effect and mechanism of herba artemisiae and licorice mixture on mice model of intrahepatic cholestasis (IC)induced by alpha-naphthylisothiocyanate (ANIT). Methods C57BL/6 female mice were randomly divided into normal group,model group,herba artemisiae group,licorice group and herba artemisiae/licorice mixture group,which were administrated with normal saline (0.2 mL/20 g),herba artemisiae (1.13 g·kg-1 · d-1 ),licorice(1 .13 g·kg-1 ·d-1 ),herba artemisiae and licorice mixture (herba artemisiae 1 .13 g·kg-1 ·d-1 and licorice 1 .13 g·kg-1 ·d-1 )for 10 days,respectively. On day 7,all groups except normal group were additionally administrated with ANIT (100 mg/kg). Serum levels of total bilirubin (TBil),alanine aminotransferase (ALT),aspartate transaminase (AST),alkaline phosphatase (ALP)and total bile acid (TBA)were detected on day 10. Hepatic pathological examinations were observed by hematoxylin and eosin staining (HE). Pregnane X receptor (Pxr),constitutive androstane receptor (Car) and their target genes were quantitated by reverse transcription-polymerase chain reaction (RT-PCR). Results Compared with those in model group,levels of TBil,ALT,AST,ALP and TBA in herba artemisiae/licorice mixture group were reduced by 22% ,23% ,25% ,20% and 30% ,respectively. Meanwhile,transcription levels of Pxr,Car,Ntcp,Oatp1 , Bsep,Mdr2,Mrp2,Mrp3,Mrp4,Cyp3a11,Ugt1a1 and Sult2a1 were elevated (P<0. 01 ). Conclusion Herba artemisiae/icorice mixture might have a certain protective effect on mice model of ANIT-induced IC. Mechanistically,it could activate nuclear receptors Pxr and Car,induce drug metabolism enzymes (Cyp3a11,Ugt1a1 and Sult2a1 )and upregulate transporters expression (Bsep,Mrp2,Mrp3,Mrp4),which promotes intrahepatic bile acid metabolism and reduces intrahepatic bile acids load.