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. This work was supported by the Sichuan Province Science & Technology Program (2022NSFSC0709), the Southwest Medical University Project (2023QN119), and the Scientific Research Project of the Luzhou Municipal Government-Southwest Medical University (2025LZXNYDYG05).

Xingqu Chen and Li Yang have contributed equally to this work. Department of Anesthesiology, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, Sichuan, China Department of Anesthesiology, The Second People’s Hospital of Yibin, Yibin, 644000, Sichuan, China Department of Pain, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, Sichuan, China Correspondence to Deshui Yu or Xiaobin Wang. The authors declare no competing interests.

This study was conducted following approval by the Medical Ethics Committee of The Second People’s Hospital of Yibin (Approval No. 2021-101-01). All animal experiments were reviewed and approved by the Laboratory Animal Center of Southwest Medical University (Approval No. 20230410-004). Procedures involving animals were performed in strict accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH Publication, 1996).