Cellular and Molecular Neurobiology

, Volume 33, Issue 1, pp 19–30

Chronic Inflammation Alters Production and Release of Glutathione and Related Thiols in Human U373 Astroglial Cells

  • Megan L. Steele
  • Stacey Fuller
  • Annette E. Maczurek
  • Cindy Kersaitis
  • Lezanne Ooi
  • Gerald Münch
Original Research

Abstract

Neurons rely on glutathione (GSH) and its degradation product cysteinylglycine released by astrocytes to maintain their antioxidant defences. This is particularly important under conditions of inflammation and oxidative stress, as observed in many neurodegenerative diseases including Alzheimer’s disease (AD). The effects of inflammatory activation on intracellular GSH content and the extracellular thiol profile (including cysteinylglycine and homocysteine) of astrocytes were investigated. U373 astroglial cells exposed to IL-1β and TNF-α for up to 96 h showed a dose-dependent increase in IL-6 release, indicative of increasing pro-inflammatory cellular activation. With increasing concentrations of IL-1β and TNF-α (0.01–1 ng/ml), an increase in both intracellular and extracellular GSH levels was observed, followed by a return to control levels in response to higher concentrations of IL-1β and TNF-α. Extracellular levels of cysteinylglycine decreased in response to all concentrations of IL-1β and TNF-α. In contrast, levels of the neurotoxic thiol homocysteine increased in a dose-dependent manner to IL-1β and TNF-α-induced activation. Our results suggest that chronically activated astrocytes in the brain might fail to adequately maintain GSH substrate delivery to neurons, thus promoting neuronal vulnerability. They might also explain the elevated levels of homocysteine found in the brains and serum of patients with AD.

Keywords

Astrocyte Glutathione Cysteinylglycine Homocysteine Inflammation Alzheimer’s disease 

Abbreviations

ABD-F

4-Flouro-7-aminosulfonylbenzofurazan

AD

Alzheimer’s disease

ANOVA

Analysis of variance

ARE

Antioxidant response element

CSF

Cerebral spinal fluid

DMEM

Dulbecco’s modified eagle medium

DTNB

5,5′-Dithio-bis(2-nitrobenzoic acid)

EDTA

Ethylenediaminetetraacetic acid

ELISA

Enzyme-linked immunosorbent sandwich assay

FBS

Foetal bovine serum

GCL

Glutamate cysteine ligase

GSH

Glutathione

GSSG

Glutathione disulfide

HPLC

High performance liquid chromatography

IL-1β

Interleukin-1 beta

IL-6

Interleukin-6

Keap1

Kelch like-ECH-associated protein 1

MRP1

Multidrug resistance protein 1

NADPH

Reduced nicotinamide adenine dinucleotide phosphate

NMDA

N-Methyl-D-aspartate

Nrf2

Nuclear factor erythroid-2-related factor 2

PD

Parkinson’s disease

ROS

Reactive oxygen species

TNB

5-Thio-2-nitrobenzoic acid

TNF-α

Tumour necrosis factor-alpha

γ-GT

γ-Glutamyltranspeptidase

γ-GCL-C

Catalytic glutamate cysteine ligase subunit

γGCL-M

Modulatory glutamate cysteine ligase subunit

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Megan L. Steele
    • 1
    • 2
  • Stacey Fuller
    • 1
  • Annette E. Maczurek
    • 1
  • Cindy Kersaitis
    • 3
  • Lezanne Ooi
    • 1
  • Gerald Münch
    • 1
    • 4
  1. 1.Department of Pharmacology, School of MedicineUniversity of Western SydneyCampbelltownAustralia
  2. 2.School of Biochemistry and Molecular BiologyJames Cook UniversityTownsvilleAustralia
  3. 3.School of Science & HealthUniversity of Western SydneyCampbelltownAustralia
  4. 4.Molecular Medicine Research GroupUniversity of Western SydneyCampbelltownAustralia

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