A systemic combined nontargeted and targeted LC-MS based metabolomic strategy of plasma and liver on pathology exploration of alpha-naphthylisothiocyanate induced cholestatic liver injury in mice

The pathology of cholestatic liver injury (CLI) was complicated, which has limited the development of anti-cholestatic drugs for a long period. Metabolomic researches focused on global and dynamic changes of the organism could shed some light on mechanism investigation. In order to characterize and validate metabolite alterations of alpha-naphthylisothiocyanate (ANIT) induced CLI in C57BL/6 mice, a systemic metabolomic approach combining nontargeted HPLC-ESI-QTOF-MS and targeted UFLC-ESI-MS/MS technologies were developed innovatively. Multivariate data analysis was applied to determine the changes of metabolites in processed plasma and liver samples between control and model groups. Afterwards, 38 potential plasma biomarkers and 17 potential liver biomarkers involved in bile acid (BA) biosynthesis, phospholipid biosynthesis, sphingolipid metabolism, alpha linolenic acid and linoleic acid metabolism, as well as arachidonic acid metabolism were found and attributed as potential biomarkers and influential pathways of cholestasis. Based on correlation analysis, BA biosynthesis played the most important role in ANIT induced CLI, thereinto, major BAs were carried out with quantitative analysis. Targeted metabolomic results showed that the increase of BAs might have an impact on intestinal microbial ecology which could aggravate liver injury probably, among which cholic acid (CA) and taurocholic acid (TCA) were the most sensitive indicators of ANIT induced CLI in both plasma and liver. In conclusion, CLI might correlate significantly with hepatocyte necrosis, metabolic disorders and imbalance of intestinal microbiome ecology triggered by BA accumulation.