Multiple protective activities of neuroglobin in cultured neuronal cells exposed to hypoxia re-oxygenation injury

Thi Thuy Hong Duong, Paul Kenneth Witting, Shane Tony Antao, Sarah Nicole Parry, Marina Kennerson, Barry Lai, Stefan Vogt, Peter Andrew Lay, Hugh Hamlyn Harris

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Abstract

Oxidative stress is associated with the pathology of acute and chronic neurodegenerative disease. We have cloned a human neuroglobin (Nb) construct and over-expressed this protein in cultured human neuronal cells to assess whether Nb ameliorates the cellular response to experimental hypoxia-reoxygenation (H/R) injury. Parental cells transfected with a blank (pDEST40) vector responded to H/R injury with a significant decrease in cellular ATP at 5 and 24 h after insult. This was coupled with increases in the cytosolic Ca2+, and the transition metals iron (Fe), copper (Cu), and zinc (Zn) within the cell body, as monitored simultaneously using X-ray fluorescence microprobe imaging. Parental cell viability decreased over the same time period with a ∼4 to 5-fold increase in cell death (maximum ∼25%) matched by an increase in caspase 3/7 activation (peaking at a 15-fold increase after 24 h) and condensation of β-actin along axonal processes. Over-expression of Nb inhibited ATP loss and except for significant decreases in the sulfur (S), chlorine (Cl), potassium (K) and Ca2+ contents, maintained cellular ion homeostasis after H/R insult. This resulted in increased cell viability, significantly diminished caspase activation and maintenance of the β-actin cytoskeletal structure and receptor-mediated endocytosis. These data indicate that bolstering the cellular content of Nb inhibits neuronal cell dysfunction promoted by H/R insult through multiple protective actions including: (i) maintenance of cellular bioenergetics; (ii) inhibition of Ca2+ influx; (iii) a reduction in cellular uptake of Fe, Cu and Zn at the expense of S, Cl and K; and (iv) an enhancement of cell viability through inhibiting necrosis and apoptosis.

Original languageEnglish
Pages (from-to)1143-1154
Number of pages12
JournalJournal of Neurochemistry
Volume108
Issue number5
DOIs
Publication statusPublished - Mar 2009
Externally publishedYes

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Keywords

  • antioxidant
  • apoptosis
  • neuro-protection
  • neuroglobin
  • oxidative stress
  • synchrotron radiation
  • x-ray fluorescence imaging

Cite this

Duong, T. T. H., Witting, P. K., Antao, S. T., Parry, S. N., Kennerson, M., Lai, B., ... Harris, H. H. (2009). Multiple protective activities of neuroglobin in cultured neuronal cells exposed to hypoxia re-oxygenation injury. Journal of Neurochemistry, 108(5), 1143-1154. https://doi.org/10.1111/j.1471-4159.2008.05846.x