Sanwubaisan Mediating Immune Response To Liver Cancer By Regulating Gut Microbiota

Sanwubaisan was first recorded in Treatise on Febrile Diseases.It is a prescription for treating cold repletion chest bind formation,and is also a representative prescription for warming bowels and purging bowels.Previous studies have confirmed that Sanwubaisan can reverse Thl/Th2 drift,and then positively regulate the immune response of liver cancer.In recent years,studies have found that the occurrence and development of liver cancer is closely related to the imbalance of gut microbiota.Sanwubaisan,as a representative prescription of warming bowels and purging bowels,has a certainly"purging bowels" effect,which may have certainly effects on the gut microbiota.Its role in anti-liver cancer immune response may be related to its intervention in the gut microbiota.Objective:To see the change of intestinal bacteria,the difference in Thl/Th2 cytokine levels,and the changes of liver cancer indicators in the intervention of Sanwubaisan on orthotopic transplantation model mice of liver cancer.Then exploring the underlying mechanism of Sanwubaisan positively regulates the immune response of liver cancer,and the further scientific connotation of the theory of "the six fu-viscera function well when unobstructed" in Chinese medicine.Method:(1)Modeling and grouping:After 46 male ICR mice of 4 week-old were adaptively reared for one week,40 mice were randomly selected for modeling orthotopic transplantation of liver cancer.After successful modeling,they were randomly divided into model group,Sanwubaisan group,vancomycin group and cholestyramine group,10 mice of each group,and the remaining 6 healthy mice were blank control group.(2)Dosing;From the second day after successful modeling,mice in the model and the blank group were given saline by gavage,and the Sanwubaisan group was administrated with Sanwubai powder suspension by gavage.Gavage was performed at a dose of 0.1 mL/10 g(weight)per day.The vancomycin group mice freely drink a vancomycin solution with a concentration of 0.5 g/L,and the cholestyramine group mice freely consume a feed containing 2%cholestyramine.(3)General condition records:The weight changes of the mice were recorded on the day of modeling and every 2 days after modeling,and the basic conditions of the mice's eating,fur color,activity status,and abdominal state were observed.Dead mice were dissected to observe the liver,tumor,ascites and so on,and the death date also was recorded.(4)The observation index:Counting the survival of mice and the occurrence of ascites after 21 days of dosing according to the above administration method,and calculating the survival rate and ascites rate of mice in each group.The following experiments were performed:liver cancer pathological tissue sections,Thl/Th2 cytokines in mouse serum and liver tissue,FXR expression in mouse ileum tissue,gut microbiota in the feces of mice and liver bile acid components and Content detection.Results:Sanwubaisan can promote Thl cytokine expression and inhibit Th2 cytokine expression.Thl and Th2 cytokine expression are related to FXR expression.In the experimental group with lower FXR expression,Thl cytokine expression is higher and Th2 cytokine expression is lower.On the contrary,the result is the opposite.Bile acid metabolism could be mediated by gut microbiota,Clostridium can convert primary bile acids into secondary bile acids in the intestine.The intestinal flora richness and diversity of the Sanwubaisan group did not change significantly according to the comparison with the model group.But he expression of Clostridium decreased,however the expression of primary bile acid increased,and the expression of secondary bile acid decreased.Bile acid could regulate FXR expression.the expression of FXR in the Sanwubaisan group reduced.Conclusion:(1)Sanwubaisan can change the structure of intestinal flora.(2)Gut microbiota can regulate bile acid metabolism.(3)Metabolism of bile acids can regulate the expression of FXR.(4)FXR expression is inversely related to the positive regulation of liver cancer immune response.(5)Sanwubaisan could regulate changes in bile acid metabolism by regulating gut microbiota,inhibit FXR expression,and reverse Thl/Th2 drift to exert positive regulatory functions of liver cancer immune response.