Study On The Effect And Mechanism Of Compound Preparations Of Penthorum Chinense Pursh On Cholestatic Liver Disease

Objective: To explore the effect and mechanism of Compound preparations of Penthorum chinense Pursh(PCP)on rats with cholestatic liver injury induced by alphanaphthylisothiocyanate(ANIT).Method: The dosage of ANIT model agent was selected.SD rats were randomly divided into 4 groups with 10 in each group,namely the blank group,the model group 1,2,3(ANIT,50,60,75 mg/kg).After 48 hours of modeling,the animals were anesthetized with chloral hydrate(0.3 mL/100 g),and the serum and liver tissues were collected.Total bilirubin(TBIL)in serum were measured,and liver tissue pathological sections were prepared to observe the histopathology changes.The pharmacodynamics study of PCP.SD rats were randomly divided into 6 groups of 10 in each group,including the blank group,the model group,the ursodeoxycholic acid(UDCA)group,and the PCP low-dose,medium-dose and high-dose groups(0.31,0.62,0.94g/kg).The experimental animals were given anesthesia 48 hours after model established.Serum and liver tissues were collected and stored for backup.Serum content of TBIL,direct bilirubin(DBIL),total bile acid(TBA),alkaline phosphatase(ALP),alanine aminotransferase(ALT),aspartate aminotransferase(AST)were measured in each group.And liver tissue pathological sections were prepared to observe the histopathological changes.Exploring the mechanism of action of PCP.The related mRNA expression in the liver tissue were detected by Quantitative Real-time PCR(RTPCR),including sodium taurocholate cotransporting polypeptide(Ntcp),bile salt export pump(Bsep),multidrug resistance-related proteins 2,3(Mrp2,Mrp3)and multidrug resistance protein1 a,1b(Mdr1a,Mdr1b).The expression of Mrp2,Mrp3 and Mdr1 b protein in liver tissue were detected by Western Blot.Result: Study on establishment of animal model of cholestatic liver injury.The results of serological indicators showed that compared with the blank group,the serum TBIL content in the model groups were obviously increased(P<0.01);compared with the model groups 2 and 3,the model group 1 serum TBIL content is lower and more discrete.Pathological changes in liver tissue: compared with the blank group,the model group 1 showed a small amount of inflammatory cell infiltration and clear outline of intrahepatic bile duct area;the model groups 2 and 3 showed a large number of inflammatory cell infiltration and degeneration and necrosis of bile duct epithelial cells.Study on Pharmacodynamics of the PCP.The results of serological indicators showed that the serum content of TBIL,DBIL,TBA and enzyme activity of ALP,ALT,AST in the model group were significantly higher than in the blank group(P<0.01).Compared with the model group,the serum content of TBIL,DBIL and enzyme activities of ALP,ALT and AST in the UDCA group and the PCP groups were obviously lower(P<0.01 or 0.05).The serum content of TBA in the UDCA group and the PCP middle and high dose groups were obviously lower(P<0.01 or 0.05),while in the PCP low dose group with no statistical significance(P>0.05).Pathological changes in liver tissue: compared with the blank group,the model group showed a large number of inflammatory cell infiltration,and hepatocytes and intrahepatic bile duct epithelial cells showed degeneration or necrosis.Compared with the model group,the UDCA group and the PCP groups showed that the above situation were significantly reduced,and histopathological damage was significantly improved.And the improvement degree in the PCP groups showed a certain dose-dependent.Study on the Mechanism of the PCP.RT-PCR results showed that compared with the blank group,the relative expression content of Ntcp and Bsep mRNA in liver tissue of the model group were significantly lower(P<0.01),and the relative expression content of Mrp2,Mrp3,Mdr1 a and Mdr1 b mRNA in liver tissue were obviously upregulated(P<0.01 or 0.05).Compared with the model group,the relative expression content of Mrp2,Mrp3,and Mdr1 b mRNA in the liver tissue of the PCP groups were obviously up-regulated(P<0.01 or 0.05),and showed a certain dose-dependent manner.Study on the Mechanism of the PCP.Western Blot results showed that compared with the blank group,the expression content of Mrp2,Mrp3,and Mdr1 b proteins in the liver tissue of the model group were obviously up-regulated(P<0.01 or 0.05).The expression content of Mrp2,Mrp3,and Mdr1 b protein in the liver tissue of the PCP groups were obviously up-regulated than those in the model group(P<0.01 or 0.05),and showed a certain dose-dependent manner.The changes in protein expression content of Mrp2,Mrp3,and Mdr1 b in liver tissues of each group were consistent with the trend of mRNA relative expression content at the gene level.Conclusion: The PCP has obvious protective effect on ANIT-induced cholestatic liver injury in rats.The mechanism of action may be through up-regulate the expression of Mrp2,Mrp3,Mdr1 b,promoting the transport of bilirubin and excretion of bile acids,and accelerating the efflux of hepatotoxic substances.In order to achieve the efficacy of curing jaundice,gallbladder and detoxification.