Preliminary Study On The Therapeutic Effect And Mechanism Of Hydrocotyle Sibthorpioides Lam.and Quercetin To Rat Cholestasis Induced By Estrogen

AimsTo investigate the therapeutic effect and the underlying mechanism of Hydrocotyle sibthorpioides Lam.and its active ingredient quercetin on e17α-Ethynylestradiol-induced cholestasis in rat.MethodsA total of 42 male Wistar rats were randomly divided into 7 groups.Except control group,the rats in the rest six groups were subcutaneously treated with e17α-Ethynylestradiol(EE,2mg/kg/d)for successive 14 days to induce chronic cholestasis.Meanwhile,rats were orally administered with saline,extract of 1-fold(1.03 g/kg/d,crude herb 10.3 g/kg/d),3-fold(3.09g/kg/d,crude herb 30.09 g/kg/d)or 9-fold(9.27 g/kg/d,crude herb 92.7g/kg/d)of human equivalent dose of Hydrocotyle sibthorpioides Lam as well as quercetin(55.62 mg/kg/d)and UDCA(60 mg/kg/d),respectively.Therapeutic effects were evaluated via comparing bile flow velocity,bile flow volume,serum biochemical indicator,hepatic pathology,as well as bile acids concentrations in serum,bile,urine,liver and kidney.Protein expression was further identified by Western Blot technique to investigate the therapeutic mechanism.ResultsBile flow experiment: Compared to the control group,the bile flow in rats of EE-treated group was significantly decreased(P<0.01).Compared with model group,the bile flow in THS9 group,quercetin group and UDCA group were all markedly increased(P<0.05),but no significant difference was observed in THS1 group and THS3 group(P>0.05).Serum biochemical indexes: Compared with normal group,the TBA and TBIL was significantly increased in model group(P<0.05).Compared with model group,serum TBA was all significantly decreased in THS1 group,THS3 group,THS9 group,quercetin group and UDCAgroup,especially in THS9 group,quercetin group and UDCA group;while TBIL was significantly decreased in THS9 group,quercetin group and UDCA group,but their level were still higher than normal group(P<0.05);the ALT was markedly reduced both in THS1 and THS9 groups(P <0.05);AST in THS9 group was significantly increased(P<0.05),no significant difference of ALP was found in all groups(P>0.05).Concentrations of bile acids: Compared with normal rats,the concentrations of TCDCA,CDCA and LCA in serum,bile and liver were all obviously increased in model rats(P<0.05).Compared with model rats,TCDCA and CDCA of serum and liver were markedly reduced(P<0.05),but TCDCA and CDCA in bile was markedly increased in THS9-treated rats,quercetin–treated rats and UDCA-treated rats(P<0.05).TCDCA,CDCA and LCA of urine and kidney in model rats were higher than that of normal rats(P<0.05).Moreover,compared with model rats,TCDCA,CDCA and LCA in kidney and urine were all significantly increased both in THS9-treated rats and quercetin-treated rats(P<0.05),TCDCA and CDCA of kidney and urine were increased in UDCA-treated group(P<0.05).No significant difference of DCA was observed in serum,bile,urine,liver and kidney in all the groups(P>0.05).4.Hepatic pathology: Compared to the normal rats,bile duct hyperplasia,venous congestion,partial hepatocytes necrosis,and inflammatory cell infiltration were observed in the liver of cholestasis model rats.The hepatic pathologic alteration in THS1 group and THS3 group was as same as that of model rats,but the alteration in THS9 group,quercetin group and UDCA group was alleviated,and the tissue structure was nearly returned to the normal condition.5.Western Blot: Compared with normal rats,the expression of hepatic Bsep,Ntcp,Mrp2,Mrp4,Oatp2 and FXR as well as Mrp4 and Oat3 of kidney in model rats were all significantly reduced(P<0.05).Compared with model rats,the expression of hepatic Bsep,Ntcp,Mrp2,Mrp4 and FXR as well as Mrp4 and FXR of kidney in THS9 group,quercetin group and UDCA group were all significantly increased(P<0.05).Compared with model rats,the expression of Oat3 in kidney was markedly increased,but the hepatic expression of Oatp2 were all reduced in THS9 group,quercetin group and UDCA group(P<0.05).Compared with Model group,only the hepatic expression of Mrp2 was increased in THS1 group,while in THS3 group,only the expression of hepatic Ntcp and FXR as well as Mrp4 in kidney was higher than that of Model group(P<0.05).ConclusionHydrocotyle sibthorpioides Lam.and its active ingredient quercetin had therapeutic effect onEE-induced cholestasis in rats,and the underling mechanism might be associated with the up-regulated expressions of hepatic Bsep,Ntcp,Mrp2,Mrp4,Oatp2 and FXR as well as Mrp4 and Oat3 in kidney.The therapeutic effect and mechanism of Hydrocotyle sibthorpioides Lam.and quercetin were as same as that of UDCA.