Exploration Of Lidan Mixture On FXR-associated Pathways In Infantile Cholestatic Liver Disease

Objective: To investigate the effect of Lidan Mixture on the mRNA and protein expression of FXR,SHP,UGT2B4 and BSEP in young rats with acute intrahepatic cholestasis induced by ?-Naphthalene isothiocyanate(ANIT)from the perspective of feedback regulation of bile acid itself.To observe the serum biochemical indexes and pathological changes of liver tissues in rats and clarify the role of Lidan Mixture in the treatment of infantile cholestatic liver disease.Methods:1.Modeling:According to the establishment of an animal model of cholestatic hepatitis,thirty weaned and young SPF level SD rats,half male and female,weighed 70-90 g,were kept in the SPF barrier system.Except the normal group,the others were administered by gavage with different drugs and were administered by gavage with ANIT at once on the fifth day,to establish the intrahepatic cholestasis model.2.Grouping and intervention: Thirty weaned and young SPF level SD rats,half male and female,weighed 70-90 g,were weighed after 3 days of adaptive feeding.Then the rats were randomly divided into normal group,model group,ursodeoxycholic acid group,Lidan Mixture high dose group and Lidan Mixture low dose group according to body weight.Normal group(N): Administered by gavage with normal saline for 6 days continuously.Model group(M): Administered by gavage with normal saline for 4 days continuously and with ANIT solution(1 ml/100 g)at once on the fifth day.Ursodeoxycholic acid(UDCA): Administered by gavage with 0.3% ursodeoxycholic acid(2ml/100g)for 6 days continuously,and given by gavage ANIT solution(1ml/100g)at intervals of 4 to 6 hours on the 5th day.Lidan Mixture high dose group(LDM-H): Administered by gavage with Lidan Mixture(7.4g/2ml)for 6 days continuously,and given by gavage ANIT solution(1ml/100g)at intervals of 4 to 6 hours on the 5th day.Lidan Mixture low dose group(LDM-L): Administered by gavage with Lidan Mixture(7.4g/4ml)for 6 days continuously,and given by gavage ANIT solution(1ml/100g)at intervals of 4 to 6 hours on the 5th day.3.Indicatrix and methods:(1)Analyze TBIL,DBIL,TBA,ALT,AST,?-GT and ALP in rats serum by automatic biochemical analyzer.(2)The hepatic hematoxylin-eosin staining of rats in each group to observe the morphological changes of the liver tissue by microscopy.(3)Real-time PCR detects the mRNA levels of FXR,SHP,UGT2B4 and BSEP.(4)Western blotting detects the protein expression levels of rat liver FXR,SHP,UGT2B4 and BSEP.Result:1.The Lidan Mixture high dose group could improve the symptoms of intrahepatic cholestasis,reduce the levels of TBIL,DBIL,TBA,ALT,AST,?-GT and ALP better than ursodeoxycholic acid group.Which also could reduce the hepatic pathological damage caused by cholestasis and repair damaged liver tissue,and also exerted a notable effect on gallbladder and liver protection.2.The m RNA expression levels of FXR,SHP,UGT2B4 and BSEP were significantly higher when compared Lidan Mixture high dose group,Lidan Mixture low dose group and ursodeoxycholic acid group with the model group;which were also higher when compared Lidan Mixture high dose group and Lidan Mixture low dose group with ursodeoxycholic acid group;In addition,compared with Lidan Mixture low dose group,the Lidan Mixture high dose group exerted significantly increase in the mRNA expression levels of FXR,SHP,UGT2B4 and BSEP.3.The protein expression levels of FXR,SHP,UGT2B4 and BSEP in the Lidan Mixture high dose group was higher than that in the ursodeoxycholic acid group.There had no obvious differences between the Lidan mixture low dose group and ursodeoxycholic acid group in the protein levels of FXR,SHP,UGT2B4 and BSEP.And the protein expression level of SHP and UGT2B4 in the Lidan Mixture high dose group was significantly higher than that in the Lidan Mixture low dose group.Conclusion:Lidan Mixture has an effect on the protection of liver and gallbladder in the young rats with acute intrahepatic cholestasis induced by ANIT,and has obvious curative impact on cholestasis and liver damage caused by it.The mechanism of Lidan Mixture can interfere FXR to promote the mRNA and protein expression levels of SHP,UGT2B4 and BSEP,promoting the excretion of bile acids in the liver and repairing the liver damage caused by cholestasis.