1. Department of Anesthesiology, Shizuoka Cancer Center, Japan.
2. Cancer Diagnostics Division, Shizuoka Cancer Center Research Institute, Japan.
4. Department of Anesthesiology, Keio University School of Medicine, Japan.
Background: The neurotoxic effects of general anesthetics may adversely affect the developing brain of young animals. Given that levels of reactive oxygen species (ROS) increase in injured mitochondria, the induction of neurotoxicity is likely mediated by oxidative stress. To evaluate this possibility, we conducted metabolomic analyses to identify the metabolites involved in anesthetic-induced neurotoxicity by exposing a well-characterized rat adrenal pheochromocytoma cell line (PC12) to isoflurane.
Methods: PC12 cells were incubated at 37°C in media with serum and then incubated with or without isoflurane for 4 h. After centrifugation, the cells were lysed in 50 μL of water for metabolomics analysis using capillary electrophoresis-mass spectrometry. Metabolite levels were quantitated according to m/z ratios and migration times relative to standards.
Results: Only cells exposed to isoflurane underwent apoptosis. We detected 102 metabolites and identified 91 in isoflurane-exposed cells and control cells. Exposure to isoflurane decreased the levels of reduced glutathione and increased those of oxidized glutathione, indicating that isoflurane accelerated oxidative stress.
Conclusions: These results suggest that oxidative stress mediates isoflurane-induced apoptosis of PC12 cells.
Keywords: Metabolomics, oxidative stress, apoptosis, isoflurane, PC12 cells