Oxidative stress, metabolomics profiling, and mechanism of local anesthetic induced cell death in yeast

Autor: Cory H.T. Boone, Dana Adamcova, Javier Seravalli, Jiri Adamec, Ryan Grove
Rok vydání: 2017
Předmět:
0301 basic medicine
Metabolomics profiling
Clinical Biochemistry
Saccharomyces cerevisiae
Oxidative phosphorylation
Biology
Pentose phosphate pathway
medicine.disease_cause
Biochemistry
03 medical and health sciences
chemistry.chemical_compound
Adenosine Triphosphate
Apoptotic cell death pathways
Tandem Mass Spectrometry
medicine
Humans
Metabolomics
Citrate synthase
Glycolysis
Flow cytometry
Anesthetics
Local

Local anesthetic toxicity
lcsh:QH301-705.5
lcsh:R5-920
Microbial Viability
Mass spectrometry
030102 biochemistry & molecular biology
Organic Chemistry
Lidocaine
Glutathione
Mitochondria
Cell biology
Adenosine Diphosphate
Citric acid cycle
Oxidative Stress
030104 developmental biology
lcsh:Biology (General)
chemistry
biology.protein
Anaplerotic reactions
Energy Metabolism
Reactive Oxygen Species
lcsh:Medicine (General)
Oxidative stress
Research Paper
Zdroj: Redox Biology, Vol 12, Iss, Pp 139-149 (2017)
Redox Biology
ISSN: 2213-2317
DOI: 10.1016/j.redox.2017.01.025
Popis: The World Health Organization designates lidocaine as an essential medicine in healthcare, greatly increasing the probability of human exposure. Its use has been associated with ROS generation and neurotoxicity. Physiological and metabolomic alterations, and genetics leading to the clinically observed adverse effects have not been temporally characterized. To study alterations that may lead to these undesirable effects, Saccharomyces cerevisiae grown on aerobic carbon sources to stationary phase was assessed over 6 h. Exposure of an LC50 dose of lidocaine, increased mitochondrial depolarization and ROS/RNS generation assessed using JC-1, ROS/RNS specific probes, and FACS. Intracellular calcium also increased, assessed by ICP-MS. Measurement of the relative ATP and ADP concentrations indicates an initial 3-fold depletion of ATP suggesting an alteration in the ATP:ADP ratio. At the 6 h time point the lidocaine exposed population contained ATP concentrations roughly 85% that of the negative control suggesting the surviving population adapted its metabolic pathways to, at least partially restore cellular bioenergetics. Metabolite analysis indicates an increase of intermediates in the pentose phosphate pathway, the preparatory phase of glycolysis, and NADPH. Oxidative stress produced by lidocaine exposure targets aconitase decreasing its activity with an observed decrease in isocitrate and an increase citrate. Similarly, increases in α-ketoglutarate, malate, and oxaloacetate imply activation of anaplerotic reactions. Antioxidant molecule glutathione and its precursor amino acids, cysteine and glutamate were greatly increased at later time points. Phosphatidylserine externalization suggestive of early phase apoptosis was also observed. Genetic studies using metacaspase null strains showed resistance to lidocaine induced cell death. These data suggest lidocaine induces perpetual mitochondrial depolarization, ROS/RNS generation along with increased glutathione to combat the oxidative cellular environment, glycolytic to PPP cycling of carbon generating NADPH, obstruction of carbon flow through the TCA cycle, decreased ATP generation, and metacaspase dependent apoptotic cell death.
Graphical abstract
Databáze: OpenAIRE