ToF-SIMS Reveals Metformin-Driven Restoration of Hepatic Lipid and Amino Acid Profiles in a Type 2 Diabetes Rat Model.

Diabetes mellitus profoundly disturbs hepatic metabolism by impairing lipid and amino acid homeostasis, yet spatially resolved molecular evidence of these alterations remains limited. This study employed Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) to visualise and quantify metabolic remodelling in rat liver under diabetic conditions and following metformin treatment. Liver cryosections from lean controls (LEAN), diabetic rats (P1), and metformin-treated diabetic rats (P2) were analysed in the negative ion mode, and all spectra were normalised to total ion counts. One-way ANOVA with false discovery rate (FDR) correction identified 43 lipid-related and 20 amino acid-related ions with significant group differences. Diabetic livers exhibited a marked depletion of phospholipid- and fatty acid-related ions (e.g., m/z 241.04, 281.25, 536.38) accompanied by increased ceramide fragments (m/z 805-806), indicating lipotoxic remodelling and mitochondrial stress. Simultaneously, aromatic and neutral amino acids such as phenylalanine, tyrosine, and glutamine were reduced, while small acidic fragments were elevated, consistent with enhanced proteolysis and gluconeogenic flux. Metformin administration partially restored both lipid and amino acid profiles toward the control phenotype. Hierarchical clustering and spatial ion maps revealed distinct group separation and partial normalisation of hepatic molecular patterns. These results demonstrate that ToF-SIMS provides label-free, spatially resolved insights into diabetes-induced metabolic disturbances and metformin-driven hepatoprotection.