Sepsis inhibits reduction of dehydroascorbic acid and accumulation of ascorbate in astroglial cultures: intracellular ascorbate depletion increases nitric oxide synthase induction and glutamate uptake inhibition.

Abstract

Sepsis is associated with oxidative stress and impaired glutamatergic transmission in brain. We investigated whether sepsis impairs accumulation of the antioxidant, ascorbate, and uptake of glutamate by astrocytes. Bacterial endotoxin (Escherichia coli lipopolysaccharide, LPS) and the inflammatory cytokine, interferon-gamma (IFNgamma), were applied to primary astrocyte cultures to model sepsis. In the absence of ascorbate, the combination of LPS and IFNgamma (LPS + IFNgammay) up-regulated inducible nitric oxide synthase (iNOS) and decreased the initial rate of glutamate uptake by 50% within 24 h. Cell viability and facilitated glucose transport activity were not affected at 24 h. Pre-treatment with ascorbate-2-O-phosphate increased intracellular ascorbate concentration and attenuated the induction of iNOS and inhibition of glutamate uptake caused by LPS + IFNgamma. Subsequent experiments examined the mechanisms by which cells accumulate ascorbate. LPS + IFNy decreased slightly the initial rate of uptake of ascorbate and inhibited markedly the rate with which intracellular dehydroascorbic acid (DHAA) was reduced to ascorbate. We conclude that septic insult impairs astrocytic clearance of DHAA from the extracellular fluid and decreases intracellular ascorbate concentration. Furthermore, sepsis induces iNOS and inhibits glutamate uptake by astrocytes through mechanisms that can be modulated by intracellular ascorbate. These results indicate treatments that increase intracellular ascorbate concentration may be beneficial for patients at risk for neurologic complication in sepsis.

• Authors: Jasminka Korcok, Feng Wu, Karel Tyml, Robert R Hammond, John X Wilson
• Journal: Journal of neurochemistry (J Neurochem) Vol. 81 Issue 1 Pg. 185-93 (Apr 2002) ISSN: 0022-3042 [Print] England
• PMID: 12067232 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Enzyme Inhibitors
• Glial Fibrillary Acidic Protein
• Lipopolysaccharides
• ascorbate-2-phosphate
• Glutamic Acid
• Interferon-gamma
• Nitric Oxide Synthase
• Nitric Oxide Synthase Type II
• Nos2 protein, rat
• Glucose
• Ascorbic Acid
• Dehydroascorbic Acid

Topics

• Animals
• Ascorbic Acid (analogs & derivatives, metabolism, pharmacology)
• Astrocytes (cytology, drug effects, metabolism)
• Biological Transport (drug effects)
• Cell Survival (drug effects)
• Cells, Cultured
• Dehydroascorbic Acid (metabolism)
• Enzyme Induction (drug effects, immunology)
• Enzyme Inhibitors (pharmacology)
• Extracellular Space (metabolism)
• Glial Fibrillary Acidic Protein (biosynthesis)
• Glucose (metabolism)
• Glutamic Acid (metabolism, pharmacokinetics)
• Interferon-gamma (pharmacology)
• Intracellular Fluid (metabolism)
• Lipopolysaccharides (pharmacology)
• Nitric Oxide Synthase (antagonists & inhibitors, metabolism)
• Nitric Oxide Synthase Type II
• Oxidation-Reduction (drug effects)
• Rats
• Rats, Wistar
• Sepsis (metabolism)
• Up-Regulation (drug effects, immunology)