Regulation Of Fish Oil Or Algal Oil On Bile Circadian Rhythm Metabolism In NAFLD Mice

objective:The effect of dietary fish oil rich in ?-3 polyunsaturated fatty acids(?-3PUFA)on bile composition and circadian metabolism with non-alcoholic fatty liver disease(NAFLD)mice was studied by bile metabonomics analysis.method:Male C57BL/6 mice were randomly divided into three groups and fed a normal chow diet(control group,CON),a high-fat diet(HFD group)or a HFD supplemented with fish oil(FO group)and a HFD supplemented with algal oil(AO group)for 12 weeks.At the end of the 12-week experiment,each group was sampled and processed at the interval of 12 hours early and late,ZTO(Zeitgeber time0,n=10)and ZT12(Zeitgeber time 12,n=10).The biochemical indexes in blood of mice were detected by kit,and the liver sections were stained with Hematoxylin-eosin(H&E).Determination and Analysis of Total metabolites in Bile by LC-MS,and different metabolites among groups were analysed by Multivariate statistical analysis;Analysis of differential metabolites pathway by KEGG database and Metaboanalyst;Detection of key proteins in bile Acid Metabolism by Western blotting.result:Compared with the CON group,the serum biochemical indexes of HFD group were abnormal,and liver steatosis and liver injury were revealed by HE staining in liver tissue sections,showing NAFLD.Compared with the HFD group,the above indexes were significantly improved in the FO or AO group(p<0.05).The bile content of each group at ZTO(CON,HFD,FO group)was significantly higher than that at ZT12(BCON,BHFD,BFO)(p<0.05).Metabonomics analysis showed that there were significant differences in bile composition among groups at the same time and involved multiple metabolic pathways(For example,primary bile acid biosynthesis,lipid metabolism,amino acid metabolism,etc.),circadian bile analysis showed that there were 22 circadian bile differential metabolites in CON group and 25 in HFD group and 43 in FO group and 27 in AO group,and homoserme was differential metabolites common to all groups.In addition,The circadian differential metabolites of CON and FO were homoserine,acetylcarnitine and dodecanoic acid,and the circadian differential metabolites of CON and AO were homoserine,D-ribose-5-phosphate and linoleic acid.conclusion:The bile components of CON,HFD and FO groups mice were different at ZTO and ZT12.Fish oil could adjust bile composition after NAFLD mice were fed with fish oil,which was related to the influence of ?-3PUFA rich fish oil on cholic acid metabolism,amino acid metabolism and lipid metabolism.?-3PUFA may also be involved in regulating the circadian rhythm of bile metabolism.