| Objetive: To explore the protective effect and mechanism of Tibetan medicine Ershiwuwei Songshi pills on ?-naphthalene isothiocyanate(ANIT)-induced cholestatic liver injury in rats.Methods: The ANIT dose of rat cholestatic liver injury caused by ANIT was determined,the model of cholestatic liver injury induced by ANIT in rats was established,and the pharmacodynamics of the Tibetan medicine Ershiwuwei Songshi Pill was evaluated.Serological indicators for rats ALP(Alkaline Phosphatase),ALT(Alanine aminotransferase),AST(Aspartate aminotransferase),DBIL(direct bilirubin),TBIL(total bilirubin)and TBA(total bile acid)The effects of the Tibetan medicine Ershiwuwei Songshi Pill on cholestasis-induced liver damage in rats induced by ANIT were evaluated in combination with the histopathological changes of the liver.Real-time fluorescence quantitative(RT-PCR)method was used to detect the relative expression changes of related gene mRNA in liver tissue,including Fxr,Cyp7a1,Mrp2,Mrp3,Bsep,and Ntcp.The possible mechanism of Ershiwuwei Songshi pills on cholestasis-type liver injury.On the basis of gene-level studies,the Western-blot method was used to detect changes in the expression of related proteins in liver tissues,and the protective mechanism of the Tibetan medicine Twenty-five Flavored Stone Pills on ANIT-induced cholestasis-induced liver injury was investigated.Results: The pharmacodynamic study of Tibetan medicine Ershiwuwei Songshi pills showed that the observation of physiological conditions: Compared with the model group,the UDCA(ursodeoxycholic acid)group and the Tibetan medicine Ershiwuwei Songshi pills were larger The physiological state of mice has been improved to a certain extent.Serological index results: Compared with the blank control group,the enzyme activities of ALT,AST,ALP,DBIL,TBIL and TBA in the model group were significantly increased(P<0.01);compared with the model group,the UDCA group and the The enzyme activities of ALT and ALP in serum of Ershiwuwei Songshi pills group were significantly reduced(P<0.01),and the enzyme activity of AST had no statistical significance(P>0.05);the UDCA group and the low-dose group of Ershiwuwei Songshi pills The serum TBA content was significantly reduced(P<0.01);the serum DBIL and TBIL content in the Ershiwuwei Songshi pills low-dose group was statistically significant(P <0.05).There was no statistically significant change in the contents of DBIL and TBIL in the high-dose group(P> 0.05).Pathological results of liver pathology in rats: Tibetan medicine twenty-five flavor pine stone pills can significantly improve liver pathological changes in the ANIT model group;mRNA analysis results show that Tibetan medicine Ershiwuwei Songshi pills can significantly change the expression of bile acid metabolism related genes Compared with the blank control group,the mRNA expression of Fxr,Mrp2 and Ntcp genes was significantly downregulated in the liver tissue of the model group(P <0.01),and the mRNA expression of Cyp7a1 and Mrp3 genes was significantly up-regulated(P <0.01);Compared with the model group,the mRNA expression of Shp in each administration group showed that the expression of rat liver tissues in the low,medium and high dose groups of Ershiwuwei Songshi pills was significantly up-regulated(P <0.01,P <0.05);rat liver tissue Fxr,MRNA expression of Mrp2 gene showed no statistical significance in each administration group(P> 0.05);mRNA expression of Ntcp gene in rat liver tissue showed that the expression of Ershiwuwei Songshi pills low-dose group was significantly increased(P <0.05),The mRNA expression of Cyp7a1 gene in each administration group was significantly down-regulated(P <0.05);the mRNA expression of Mrp3 gene in each administration group was significantly downregulated(P <0.01);mRNA of Bsep gene in rat liver tissue of UDCA group The expression was significantly down-regulated(P <0.05);the mRNA expression of Bsep gene in rat liver tissue showed no statistical significance in the low,medium and high dose groups of Ershiwuwei Songshi pills(P> 0.05).The protein expression results showed that,compared with the blank control group,the expression levels of Fxr and Shp proteins in the liver tissue of the model group were significantly reduced(P <0.01),and the expression levels of UGT2B4 protein were significantly increased(P <0.05).Compared with the model group,the expression levels of Fxr and Shp protein in liver tissues of each dosage group of Ershiwuwei Songshi pills were significantly increased(P <0.01,P <0.05);the expression levels of Fxr and Shp protein in liver tissues of UDCA group The change was not statistically significant(P> 0.05);UGT2B4 protein expression in liver tissue of UDCA group,Ershiwuwei Songshi pills middle-dose group and high-dose group was significantly increased(P <0.01).Conclusion: Tibetan medicine Ershiwuwei Songshi Pill has obvious protective effect on cholestasis-induced liver injury induced by ANIT in rats.The protective mechanism may be through mediating FXR signal pathway,regulating its downstream genes and proteins,and inhibiting bile acid secretion.And synthesis,make the metabolism of bile acids tend to a steady state,and then improve cholestasis.It has no obvious regulating effect on the bile acid conversion process. |
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