Metabolite alteration analysis of acetaminophen-induced liver injury using a mass microscope

Acetaminophen (APAP)-induced liver injury (APAP-ILI), which occurs during APAP overdose, has been extensively studied. The production of N-acetyl-p-benzoquinone imine (NAPQI), the reactive metabolite of APAP, primarily contributes to liver injury. However, the mechanism underlying APAP-ILI has not been fully characterized. For further clarification, it is important to consider drug localization and endogenous substances in the injured liver. Herein, we show the localization of NAPQI metabolites and the injury site-specific changes in endogenous substances in the rat liver following APAP overdose using a mass microscope. Our results of on-tissue derivatization matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) showed that the glutathione metabolite of APAP, a detoxified metabolite of NAPQI, localized in the damaged central vein region in the rat liver following APAP administration. Moreover, in the conventional MALDI-MSI, the intensities of some phospholipids, phosphocreatine, and ceramides decreased or increased in the damaged regions compared with those in non-damaged regions. Phosphocreatine was localized in the damaged cells, whereas its related mitochondrial creatine kinase was localized in the non-damaged cells. These results are expected to contribute to further elucidation of the mechanisms underlying APAP-ILI. Our findings illustrate the localization of NAPQI-related metabolites and endogenous molecules associated with APAP-ILI, which may be related to apoptosis or metabolic adaptation ultimately protecting the cells. As MALDI-MSI can analyze and differentiate regions with tissue damage, it is a valuable tool for analyzing the mechanism underlying drug-induced liver injury to identify novel biomarkers.