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

Research output: Contribution to journalArticlepeer-review

56 Citations (Scopus)

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

Keywords

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

Fingerprint Dive into the research topics of 'Multiple protective activities of neuroglobin in cultured neuronal cells exposed to hypoxia re-oxygenation injury'. Together they form a unique fingerprint.

Cite this