This study constitutes a secondary analysis of single-nucleus RNA sequencing (snRNA-seq) data with marmoset-specific metabolomic profiling derived from our prior publication (Zhang et al., Med Gas Res, 2025)，raw FASTQ files are available in GEO under accession GSE234493. The neurological effects of sevoflurane anesthesia in the aging brain remain a critical concern, yet the translatability of findings from rodent models to humans is limited by interspecies differences. To address this, we conducted a cross-species comparison of sevoflurane-induced neurotoxicity in aged mice and marmosets, integrating single-cell transcriptomics and species-specific metabolomics. Our results reveal that sevoflurane disrupts nitric oxide (NO) signaling and calcium homeostasis in both species, leading to endoplasmic reticulum stress and oxidative damage, but with distinct pathological outcomes. In mice, excitatory neurons exhibited pronounced protein misfolding, apoptotic activation, and glycolytic upregulation, whereas marmosets uniquely activated the pentose phosphate pathway (PPP) in microglia—a metabolic adaptation linked to NADPH-mediated antioxidant defense. Notably, NO-mediated neurotoxicity originated primarily from neurons in marmosets but endothelial cells in mice, highlighting fundamental differences in synaptic vulnerability. These findings demonstrate that while conserved pathways (e.g., calcium/NO dysregulation) underlie anesthesia-induced injury, primates employ evolved neuroprotective mechanisms absent in rodents. Our work underscores the importance of non-human primate models in translational anesthesia research and identifies PPP potentiation as a potential therapeutic strategy for perioperative neuroprotective interventions in elderly patients.