Metabolomic profiling reveals persistent metabolic alterations and selective working memory deficits following neonatal anesthesia in rats

Preclinical studies have established that common pediatric anesthetics such as sevoflurane and propofol can induce neurotoxicity, yet their molecular mechanisms remain incompletely elucidated. This study combined LC-MS-based metabolomics with caspase-3 and cleaved caspase-3 immunoblotting, Y-maze, and Morris water maze testing to investigate metabolic alterations linked to anesthesia-induced neurotoxicity in neonatal Sprague-Dawley rats. Animals at postnatal day 7 (P7) were exposed to a single 2-hour session of sevoflurane or propofol anesthesia and assessed at 6 h (P7) and 35 days (P42) post-exposure. Both anesthetics significantly upregulated hippocampal caspase-3 and cleaved caspase-3 expression at both time points and resulted in long-term decline of working memory in the Y-maze test at P42. However, no significant cognitive alteration was detected in the Morris water maze test at P42. Metabolomic analysis revealed anesthetic-specific metabolic disturbances in the hippocampus. In propofol-treated rats, six metabolites—indole, isoleucine, lysine, phenylalanine, D-ribose 5-phosphate, and biotin—were consistently and significantly elevated at 6 h and 35 days post-anesthesia. All six metabolites showed a strong positive correlation with caspase-3 levels. In contrast, sevoflurane anesthesia led to a marked and sustained downregulation of phosphatidylcholine PC (16:0/16:0), which correlated negatively with caspase-3 expression. This study identifies distinct hippocampal metabolite profiles associated with prolonged neurotoxicity induced by propofol and sevoflurane. The propofol-related metabolites—indole, isoleucine, lysine, phenylalanine, D-ribose 5-phosphate, and biotin—and the sevoflurane-associated metabolite PC (16:0/16:0) represent potential intervention targets for mitigating the adverse neurodevelopmental effects of general anesthesia in neonates.