Effect of Hirudin on farnesol X receptor pathway during acute intrahepatic cholestasis

Objective:The purpose of this study is to explore the effect of Hirudin on the farnesoid X receptor(FXR)pathway during acute intrahepatic cholestasis in vivo and in vitro.Method:In vivo,sixty male Sprague-Dawley rats were randomly divided into six groups:regular group,model group,ursodeoxycholic acid(UDCA)group(60 mg/kg),hirudin treatment group(84 u/kg),hirudin treatment group(63 u/kg)and hirudin treatment group(42 u/kg).The male Sprague-Dawley rats of UDCA group were intragastrically administered with a corresponding concentration of 0.005 mL/g body weight for seven days,once a day;and the hirudin treatment group was injected subcutaneously with different concentrations of Hirudin for seven days,once a day;Except for the normal group,other groups of rats were given 100 mg/kg ANIT by gavage on the 5th day.The model was administered by gavage once a day for three days.In vitro,(Z)-Guggulsterone was used to stimulate the L02 cells(0.05μmol/ml),with or without different concentrations of Hirudin(2,4 and 8 u/ml)for 24 h.The liver tissue was examined by HE microscope and the pathological state of the rat liver was observed;FXR,Small heterodimeric chaperone receptor(SHP),uridine diphosphate glucuronide transfer 2B4(UGT2B4),bile salt output pump(BSEP)mRNA and protein expressions were tested by real-time fluorescent quantitative PCR and Western blot test.And immunohistochemistry(IHC)was used to analyze the expression of FXR.Results:Compared with the model group,the hirudin group can improve liver tissue damage,and promote FXR,SHP,BSEP and UGT2B4 proteins and mRNA expression in vivo and in vitro.Conclusion:Hirudin can alleviate intrahepatic cholestasis,reduce liver tissue damage.Hirudin can up-regulate the expression of FXR gene,promote the up-regulation of SHP,BSEP and UGT2B4 genes,and inhibit the cholestasis pathway to protect liver cells.The study may provide an effective drug for clinical treatment of intrahepatic cholestasis.