Study On Mechanism Of Fermented Bile Powder In Treating Acute Alcoholic Liver Injury

AIM:The optimal fermentation days for different fermentation days were screened out through component detection.The changes of bile acid content in bile powder before and after fermentation were compared,and the role of bacteria in the fermentation process was analyzed.The pharmacological effects of fermented bile powder were analyzed by acute alcoholic liver injury model.The mechanism of fermented bile powder in the treatment of acute alcoholic liver injury was analyzed by metabolomics study.METHOD : The three strains isolated from the bezoar were mixed with bile and fermented,and 7 d,15 d,22 d,30 d,37 d and 45 d of fermentation were selected.The bile powder of different fermentation days was analyzed by UHPLC-MS mass spectrometry with the content of bile acid as the index.Choose the fermented bile powder with the highest bile acid content.UHPLC-MS mass spectrometry was used to explore the differences in bile acid components between ox bile powder and bile powder with the best fermentation days to analyze the role of bacteria in it.The pharmacological effects of fermented bile powder were analyzed through the model experiment of acute alcoholic liver injury.UHPLC-MS mass spectrometry was used to analyze the serum of mice in the blank group,model group and drug group,and orthogonal partial least squares discriminant(OPLS-DA)was used for multivariate analysis to screen differential metabolites.The differential metabolites were introduced into Metabo Analyst to construct metabolites.Pathway;UHPLC-MS mass spectrometry was used to analyze the livers of mice in the blank group,the model group and the administration group,and the orthogonal partial least squares discriminant method(OPLS-DA)was used for multivariate analysis to screen for differential metabolites.RESULTS:Using the content of cholic acid as an indicator,the bile powders fermented for 7d,15 d,22d,30 d,37d and 45 d were analyzed by UHPLC-MS mass spectrometry,and it was found that the content of cholic acid was the highest at the 30 th day of fermentation.The differences in bile acids between ox bile powder and bile powder after 30 days of fermentation were investigated by UHPLC-MS mass spectrometry.It plays a role in promoting the conversion of conjugated bile acids into free bile acids.Through the acute alcoholic liver injury model experiment,it was found that fermented bile powder can treat acute alcoholic liver injury,mainly by regulating the body weight,spleen coefficient and ALT,AST,GSH,MDA,SOD and IL-6,IL-1β,TNF-α of mice level,of which the treatment group had the best effect at high doses.Through the study of mouse serum and liver metabolomics,it was found that the main influencing pathway of fermented bile powder in the treatment of acute alcoholic liver injury is the biosynthetic pathway of primary bile acids,and the main metabolites affected are Tauro ω-Muricholic Acid、Taurohyocholic Acid、5β-Cholanic Acid-3α,6α-diol-7-one、Murocholic Acid、Taurolithocholic Acid、3α,12α,23-Nordeoxycholic Acid and Chenodeoxycholic Acid levels.CONCLUSION:The research on the bile acid composition of bile powder before and after fermentation showed that the bacteria could promote the degradation of bound bile acids into free bile acids,and play a role in enhancing the activity of bile acids.Fermented bile powder can treat acute alcoholic liver injury,mainly affecting the biosynthetic pathway of primary bile acids.It may be that bile acid components in bile powder act on bile acid receptors in the liver to cause changes in bile acid levels.