Study The Effects Of Wuzhi Capsule And Its Quality Control Ingredient,schisandrin A,on Cholestatic Liver Injury

ObjectivesCholes deposition refers to the formation and efflux of bile in hepatocytes or bile ducts,resulting in excessive accumulation of bile acids such as bile acids,cholesterol and bilirubin in the liver and systemic circulation,thereby damaging liver cells.Most drugs have clinical side effects of liver injury.Drug-induced liver injury is divided into cell-induced liver injury,cholestatic liver injury and mixed liver injury.Cholestatic liver injury is a common clinical disease in modern clinic.The pathogenesis of the disease is complicated,and the current clinical treatment of cholestatic liver damage is very limited.Clinical observations and studies have shown that wuzhi capsules can counteract the increase in transaminase and bilirubin caused by TB drugs combination and slow down liver damage caused by drugs.Based on clinical reports and previous work,we hypothesized that wuzhi capsules and schisandrin A can antagonize rifampicin-induced cholestatic liver injury.Therefore,this study used rifampicin for the modeling of cholestatic liver injury,and studied the hepatoprotective mechanism of the five ester capsules and its quality control component,Schisandrin A,in vivo and in vitro,for the clinical development of treatment of cholestatic.The drugs for liver damage provide theoretical and experimental evidence.Research methods1.Study on the effects and mechanism of Wuzhi Capsule and Schisandrin A against cholestatic liver injuryA rat model of cholestasis-induced liver injury induced by rifampicin was established and evaluated from four aspects.1 serum ALT,AST,ALP,TB,DB and TBA and other biochemical indicators of cholestatic liver injury;2 liver tissue pathology examination,rat liver pathological section evaluation;3 protein extraction,Western blotting method for determination of MRP2,BSEP and Protein expression of PXR;4 RNA extraction,real-time fluorescent quantitative PCR to determine mRNA levels of MRP2,BSEP and PXR.Three doses of pentaester capsule and schisandra A were administered by intragastric administration,and a positive control group(administered with ursodeoxycholic acid)was set up from serum biochemical indicators,pathological sections,protein expression and mRNA levels.In one aspect,the antagonistic effect of Wuzhi Capsule and Schisandrin A on cholestasis-induced liver injury induced by rifampicin was evaluated.2.Cell level study on the effects and mechanism of anti-cholestasis liver injury induced by Wuzhi capsule and its quality control component Schisandra AHepG2 cells were used as the research object,and a series of rifampicin cells were administered in vitro.The cell model of cholestatic liver injury was constructed by cell supernatants such as ALT,AST,TB,DB and other indicators of cholestatic liver injury.In the cell model of cholestatic liver injury,the effects of five ester capsules and schisandrin A on rifampicin-induced cholestatic liver were evaluated from three aspects:biochemical indicators of liver injury,related protein(MRP2,BSEP and PXR)expression and mRNA levels.The mechanism of action of the injury.The silencing cell line of PXR gene was silenced by the lentiviral interfering RNA technology of our group.Compared with the normal HepG2 cell group,the PXR pathway was explored in wuzhi capsules from three aspects:biochemical indicators of liver function,expression of related proteins and mRNA levels.And the role played by schisandrin A against rifampicin-induced cholestatic liver injury.Result1.A daily dose of 100 mg/kg of rifampicin was administered once a day,and there was a significant effect of causing cholestasis after one week of administration.Compared with the blank control group,the liver function in the serum of the model group AST,TB,DB,TBA increased significantly(P<0.01),and there was no significant change in serum ALT and ALP(P>0.05).HE staining of liver pathological sections showed that the liver tissue of the model group had obvious inflammatory cell infiltration,accompanied by bile flow and bile duct damage.Compared with the rifampicin-induced model group,high-dose wuzhi capsules and schisandrin A group significantly reduced the levels of liver injury markers such as AST,TB,DB,and TBA in rat serum(P<0.001).Liver pathology analysis also improved relative to the model group.MRP2,BSEP and PXR protein and mRNA levels were significantly decreased in the model group,and the protein and mRNA levels of the rats in each treatment group were significantly increased and dose-dependent.2.The results of cell experiments showed that rifampicin can cause obvious liver damage in normal HepG2 cells and silenced cells,and DB and TB increased significantly(P<0.01).There was no significant difference between AST and ALT,suggesting that Lifu The liver injury caused by ping is a cholestatic liver injury.After the hepatic injury cells were given to the wuzhi capsule,the expression of the protein was detected,and the PXR and related regulatory proteins were down-regulated,and the mRNA level was also decreased.And the cells treated by the pentaester capsule significantly reduced the liver biochemical indicators of the cells,and the corresponding proteins were also up-regulated.However,the administration of Schisandrin A did not differ significantly at the cellular level.ConclusionsWuzhi capsule and its quality control ingredient,Schisandrin A,have protective effects on rifampicin-induced injury.Combined with in vitro and in vivo studies,the hepatoprotective mechanism of Wuzhi Capsule may be regulated by nuclear receptor PXR.The bile acid transporter MRP2 and BSEP function to regulate bile transporter function,and its role may also be related to other nuclear receptors.