The Analysis Of The Composition Of Different Animal Bile Powders And Their Effect On Nonalcoholic Fatty Liver Disease

Objective:To compare the activity of bile in four kinds of animals:bears,pigs,cows and chickens.To construct a model of nonalcoholic fatty liver disease and to provide treatment using the bile from the four different animals.To explore the effects of different bile in by analyzing the bioactive components of each animal bile,in order to provide a theoretical basis for clinical application.Methods:1:HPLC-Q-TOF MS composition analysis:bear bile powder,pig bile powder,bile powder and chicken bile powder composition analysis,including retention time,precise molecular weight and standard comparison,and compare to existing literature to identify differences in different bile ingredients;2:Construct animal model and intervention:the animal model selected in this experiment is a high fat diet rat model,resulting in non-alcoholic fatty liver disease,which is currently internationally recognized as the most convenient,model with highest success rate and stability.The model can also be a good simulation of the occurrence and pathological process of fatty liver disease from scratch,from mild to severe.Sixty male SD rats were randomly divided into two groups:normal control group(n = 10)and control group(n = 60).The high fat diet group was divided as:model group(n = 10),bile powder intervention group(n = 10),pig bile powder intervention group(n = 10),bovine bile powder intervention group(n = 10)and chicken bile powder intervention group(n = 10).After the rats were fed with high fat diet for 6 weeks,the rats in different groups were treated with corresponding volume of normal saline for 7 weeks.3:Materials:after 24h of the last dose,the abdominal aorta was injected with a 3%pentobarbital sodium,60mg/kg,and then the abdominal aorta was executed,and the liver was taken and weighed.Separate serum-20 ℃ preservation,used for determination of blood biochemical indicators;Rapid separation of the entire liver,weighing,and then taking the right lobe of the liver tissue 1 x 1 x 1cm3,fixed with 4%paraformaldehyde(in pH 7.4 PBS),used for morphological detection.4:Recording of the general situation of rats during the experiment.Following the treatment period,the biochemical analysis of various markers in rat serum were evaluated,including:glutamate pyruvate transaminase(AST),glutamate pyruvate transaminase(ALT),glutamate pyruvate transaminase(ALT),glutamate pyruvate transaminase(ALT)and glutamate pyruvate transaminase(ALT),Triglyceride(TG),direct bilirubin(DBIL),alkaline phosphatase(ALP),lactate dehydrogenase(LDH),low density lipoprotein lipoprotein,LDL)and total cholesterol(CHOL).Changes in final weight and liver weight were recorded and HE staining was used to examine the histopathological changes of the liver.Results:Treatment with different types of animal bile powders had different effects on high-fat diet-induced increase of body weight and/or liver weight in rats.Notably,bear bile and pig bile displayed inhibitory effects on the change of both body weight and liver weight in rats;and bear bile powder had most potent activity(P weight<0.05,P liver weight<0.01).Cow bile powder showed inhibitory effect only on body weight(P<0.05)but not on liver weight(P>0.05),whereas treatment with chicken bile powder had no effect on either body weight or liver weight(P>0.05).In addition,high-fat diet induced Nonalcoholic Fatty liver disease in rats,as evidenced by the significant increased in the serum expression of various biochemical markers including glutamic oxalacetic transaminase(AST),glutamate pyruvate transaminase(ALT),triglyceride(TG),direct bilirubin(DBIL),alkaline phosphatase(ALP),lactic dehydrogenase(LDH),low-density lipoprotein(LDL)and total cholesterol(CHOL),which however could be reversed by treatment with all kinds of bile powders.In particularly,bear bile and chicken bile showed the most potent therapeutic efficacy against Nonalcoholic Fatty liver disease.Moreover,cow and bear bile effectively alleviated high-fat diet induced pathohistological damages in liver sections,as evidenced by liver enlargement and discoloration,hepatocyte swelling,infiltration of inflammatory cells and formation of lipid vacuoles,whereas pig bile and chicken bile did not show significant therapeutic effects.Furthermore,bioactive component analysis revealed that there were significant differences in the relative content of taurocholic acid,taurodeoxycholic acid and ursodeoxycholic acid among different types of animal bile.More interestingly,a unique component with molecular weight of 496.2738 Da,whose function has not yet been reported,was identified only in bear bile powder.Conclusion:This study showed that the bile from the four kinds of animals,bear,pig,cow and chicken,had different therapeutic effects on nonalcoholic fatty liver disease caused by high fat diet,and the overall effect of bear bile powder was the most significant.At the same time,different types of animal bile had different bioactive components,which may be the reason why bear,cow,pig or chicken bile had different effects on the treatment of non-alcoholic fatty liver disease.However,a compound with molecular weight of 496.2738 Da,which was only found in bear bile powder,may present a novel way for the study and treatment of non-alcoholic fatty liver disease in the future.