Dynamic Spatial Metabolomics Reveals Region-Specific Metabolic Reprogramming and Markers during Vascular Cognitive Impairment Progression.

Vascular cognitive impairment (VCI) significantly contributes to dementia; however, the precise metabolic mechanisms underlying its region-specific pathological progression remain poorly understood. We hypothesized that spatially resolved metabolomics could uncover detailed spatiotemporal metabolic disruptions and key therapeutic windows. Using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), we systematically mapped metabolite alterations in hippocampal subregions (CA1, CA3, dentate gyrus) and cortical areas of a rat model of chronic cerebral hypoperfusion (2-VO) at early (7 days), intermediate (28 days), and late (56 days) stages. MALDI-MSI enabled region-resolved visualization of lipid, energy, and inflammatory metabolic signatures at a spatial resolution of 20 μm, allowing comparative analysis of spatiotemporal metabolic trajectories across disease progression. Our analysis revealed distinct temporal patterns: early depletion of membrane lipids and cholinergic precursors primarily in CA1 and DG, midphase ceramide accumulation and glycolytic shift notably in CA3 at day 28, and widespread mitochondrial dysfunction with necroptotic signaling by day 56. Critical metabolic putative markers─including PC(15:0/16:0), CerP(d18:1/18:0), and LysoPA(0:0/18:1)─delineated disease stages, establishing day 28 as a pivotal therapeutic intervention window. These findings provide novel mechanistic insights and metabolic targets for therapeutic strategies in VCI.