Hypoxia-triggered O-GlcNAcylation in the brain drives the glutamate-glutamine cycle and reduces sensitivity to sevoflurane in mice

Br J Anaesth. 2022 Nov;129(5):703-715. doi: 10.1016/j.bja.2022.06.041. Epub 2022 Aug 26.

Abstract

Background: Hypersensitivity to general anaesthetics predicts adverse postoperative outcomes in patients. Hypoxia exerts extensive pathophysiological effects on the brain; however, whether hypoxia influences sevoflurane sensitivity and its underlying mechanisms remain poorly understood.

Methods: Mice were acclimated to hypoxia (oxygen 10% for 8 h day-1) for 28 days and anaesthetised with sevoflurane; the effective concentrations for 50% of the animals (EC50) showing loss of righting reflex (LORR) and loss of tail-pinch withdrawal response (LTWR) were determined. Positron emission tomography-computed tomography, O-glycoproteomics, seahorse analysis, carbon-13 tracing, site-specific mutagenesis, and electrophysiological techniques were performed to explore the underlying mechanisms.

Results: Compared with the control group, the hypoxia-acclimated mice required higher concentrations of sevoflurane to present LORR and LTWR (EC50LORR: 1.61 [0.03]% vs 1.46 [0.04]%, P<0.01; EC50LTWR: 2.46 [0.14]% vs 2.22 [0.06]%, P<0.01). Hypoxia-induced reduction in sevoflurane sensitivity was correlated with elevation of protein O-linked N-acetylglucosamine (O-GlcNAc) modification in brain, especially in the thalamus, and could be abolished by 6-diazo-5-oxo-l-norleucine, a glutamine fructose-6-phosphate amidotransferase inhibitor, and mimicked by thiamet-G, a selective O-GlcNAcase inhibitor. Mechanistically, O-GlcNAcylation drives de novo synthesis of glutamine from glucose in astrocytes and promotes the glutamate-glutamine cycle, partially via glycolytic flux and activation of glutamine synthetase.

Conclusions: Intermittent hypoxia exposure decreased mouse sensitivity to sevoflurane anaesthesia through enhanced O-GlcNAc-dependent modulation of the glutamate-glutamine cycle in the brain.

Keywords: anaesthesia; glucose metabolism; glutamate–glutamine cycle; glutamine synthetase; glycosylation; hypoxia; sevoflurane.

MeSH terms

  • Acetylglucosamine* / metabolism
  • Acetylglucosamine* / pharmacology
  • Anesthetics, General* / pharmacology
  • Animals
  • Brain
  • Diazooxonorleucine / pharmacology
  • Glucose / metabolism
  • Glutamate-Ammonia Ligase / metabolism
  • Glutamate-Ammonia Ligase / pharmacology
  • Glutamates / pharmacology
  • Glutamine / pharmacology
  • Hypoxia
  • Mice
  • Oxygen / pharmacology
  • Sevoflurane / pharmacology

Substances

  • Acetylglucosamine
  • Sevoflurane
  • Glutamine
  • Diazooxonorleucine
  • Glutamate-Ammonia Ligase
  • Glucose
  • Anesthetics, General
  • Oxygen
  • Glutamates