Bicyclol Attenuates Mice Liver Injury Of Cholestasis Induced By Bile Duct-Ligated

Objects Cholestasis is a kind of pathological state of bile secretion and excretion disorder.Acute cholestasis can rapidly lead to hyperbilirubinemia,hepatocyte death,fibrosis,liver failure and even death.Ursodeoxycholic acid(UDCA)is the most effective treatment for cholestasis,but it is not very kind for some patients.There were many reports shows that bicyclol can enhance the detoxification function of liver,reduce the transaminase level of patients,and improve various biochemical indexes of liver.It is a popular medicines of liver protection in clinic,which can remove oxygen free radicals,protecting the stability of liver cell membrane,mitochondria of hepatocytes and inhibiting the expression of inflammatory cytokines.The aim of this study is to observe the therapeutic effect and possible mechanism of bicyclol on cholestatic liver injury induced by ligation of common bile duct in mice.Methods There were 18 C57BL/6 healthy male mice were randomly divided into control group(sham-operation),and bile duct ligation surgery(BDL)group,and BDL+bicyclol group.24 hours after surgery,bicyclol group were randomly given the gavage of100mg/kg bicyclol(dissolved in 0.5%CMC)for 14 days,meanwhile,control group and BDL group were randomly given the gavage of the same volume of 0.5%Carboxymethyl cellulose(0.5%CMC,control and solvent)for 14 days too.Their body weighed were measured daily.After an overnight abrosia,they were randomly sacrificed and the blood was collected for serum total bilirubin,alanine aminotransferase(ALT),aspartate aminotransferase(AST),γ-glutamyl transpeptidase(GGT),alkaline phosphatas(ALP)and total bilirubin(TBIL)analyses,and liver tissue for liver histology to assess the liver injury and liver fibrosis and quantitative real-time polymerase chain reaction(PCR)and Western Blot analysis.Messenger RNA expression of farnesoid X receptor(FXR),cytochrome P450 7A1(CYP7A1),bile salt export pump(BSEP),transforming growth factor β1(TGF-β1),collagen 1a1(COL1A1),α-smooth muscle actin(α-SMA)and tumor necrosis factor α(TNF-α)and interleukin-β1(IL-1β)were detected by RT-PCR analysis.And the protein ofSmad2/3 were analyzed by Western Blot analysis.Result1.Compared with the sham-operation,BDL mice showed higher level of Serum ALT,AST,ALP,GGT and TBIL(P<0.001),however,these liver enzymes of the treatment with bicyclol were significantly reduced(P<0.05).2.HE staining of liver under microscope,the livers of sham-operation had no abnormalities.In BDL,small bile duct with dilation,proliferation,infiltration of inflammatory,degeneration and necrosis were obviously observed.But after bicyclol intervention these change were getting better.3.The m RNA expression: In the BDL mice’s livers,the m RNA expression of CYP7A1,TGF-β1,COL1A1,IL-1β and TNF-α was remarkablely increased(P <0.001),while the m RNA expression of FXR in the liver was decreased(P<0.001).Nevertheless,the m RNA expression of CYP7A1,TGF-β1,COL1A1,IL-1β and TNF-α in the liver was remarkablely decreased(P<0.05)after using bicyclol,while the m RNA expression of FXR in the liver was increased(P<0.05).4.The protein expression: In BDL,the liver protein expression of Smad2/3 was prominently increased(P<0.001),but after the gavage of bicyclol it was reversed(P<0.05).Conclusion In the model of BDL cholestatic liver injury,bicyclol shows inflammation inhibition and liver enzyme decrease.In addtion,bicyclol also attenuates cholestasis,its mechanism consist of bicyclol maybe activate FXR and BSEP but inhibit the key enzyme of bile acid synthesis(CYP7A1).Moreover,this experiment also suggests that bicyclol has a certain effect in hepatic fibrosis,and ts maybe related to TGF-β1.