Pyruvate protects the brain against ischemia-reperfusion injury by activating the erythropoietin signaling pathway

Myoung Gwi Ryou, Ran Liu, Ming Ren, Jie Sun, Robert T. Mallet, Shao Hua Yang

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Background and Purpose-Pyruvate is known to be cytoprotective through antioxidant and anti-inflammatory mechanisms. We tested the hypothesis that pyruvate protects the brain against ischemia-reperfusion injury by inducing endogenous erythropoietin (EPO) expression. Methods-Pyruvate's protective effect was evaluated in C6 glioma cells and HT22 neuronal cells subjected to transient oxygen glucose deprivation. Cell viability (calcein AM assay) and expression of hypoxia-inducible factor-1α, EPO, Akt and Erk (immunoblot), and EPO receptor (reverse transcription-polymerase chain reaction) were analyzed. Transient focal cerebral ischemia in rats was induced by 2 hours middle cerebral artery occlusion followed by 24 hours reperfusion. Pyruvate or saline was infused from 60 minutes occlusion until 30 minutes reperfusion. Lesion volume and DNA fragmentation were assessed by 2,3,5-triphenyltetrazolium staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay, respectively. Immunoblots were conducted to determine cerebral EPO contents. Results-Pyruvate increased cell viability, hypoxia-inducible factor-1α, EPO, and Akt phosphorylation. Small interfering RNA suppression of hypoxia-inducible factor-1α and EPO abolished pyruvate-induced cytoprotection. In the rat stroke model, pyruvate reduced lesion volume by 84% and DNA fragmentation by 77% versus controls; increased EPO content paralleled these cerebroprotective actions of pyruvate. Conclusions-Pyruvate activation of the hypoxia-inducible factor-1α-EPO signaling cascade in neurons and glia could protect the brain from ischemia-reperfusion injury.

Original languageEnglish
Pages (from-to)1101-1107
Number of pages7
JournalStroke
Volume43
Issue number4
DOIs
StatePublished - 1 Apr 2012

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Erythropoietin
Reperfusion Injury
Brain Ischemia
Pyruvic Acid
Hypoxia-Inducible Factor 1
DNA Fragmentation
Reperfusion
Cell Survival
Erythropoietin Receptors
Cell Hypoxia
Cytoprotection
DNA Nucleotidylexotransferase
Middle Cerebral Artery Infarction
Transient Ischemic Attack
Glioma
Neuroglia
Small Interfering RNA
Reverse Transcription
Anti-Inflammatory Agents
Antioxidants

Keywords

  • brain recovery
  • erythropoietin
  • focal ischemia
  • hypoxia-inducible factor
  • pyruvate
  • stroke care

Cite this

@article{62d97249053341d2a897f9536ed52098,
title = "Pyruvate protects the brain against ischemia-reperfusion injury by activating the erythropoietin signaling pathway",
abstract = "Background and Purpose-Pyruvate is known to be cytoprotective through antioxidant and anti-inflammatory mechanisms. We tested the hypothesis that pyruvate protects the brain against ischemia-reperfusion injury by inducing endogenous erythropoietin (EPO) expression. Methods-Pyruvate's protective effect was evaluated in C6 glioma cells and HT22 neuronal cells subjected to transient oxygen glucose deprivation. Cell viability (calcein AM assay) and expression of hypoxia-inducible factor-1α, EPO, Akt and Erk (immunoblot), and EPO receptor (reverse transcription-polymerase chain reaction) were analyzed. Transient focal cerebral ischemia in rats was induced by 2 hours middle cerebral artery occlusion followed by 24 hours reperfusion. Pyruvate or saline was infused from 60 minutes occlusion until 30 minutes reperfusion. Lesion volume and DNA fragmentation were assessed by 2,3,5-triphenyltetrazolium staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay, respectively. Immunoblots were conducted to determine cerebral EPO contents. Results-Pyruvate increased cell viability, hypoxia-inducible factor-1α, EPO, and Akt phosphorylation. Small interfering RNA suppression of hypoxia-inducible factor-1α and EPO abolished pyruvate-induced cytoprotection. In the rat stroke model, pyruvate reduced lesion volume by 84{\%} and DNA fragmentation by 77{\%} versus controls; increased EPO content paralleled these cerebroprotective actions of pyruvate. Conclusions-Pyruvate activation of the hypoxia-inducible factor-1α-EPO signaling cascade in neurons and glia could protect the brain from ischemia-reperfusion injury.",
keywords = "brain recovery, erythropoietin, focal ischemia, hypoxia-inducible factor, pyruvate, stroke care",
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Pyruvate protects the brain against ischemia-reperfusion injury by activating the erythropoietin signaling pathway. / Ryou, Myoung Gwi; Liu, Ran; Ren, Ming; Sun, Jie; Mallet, Robert T.; Yang, Shao Hua.

In: Stroke, Vol. 43, No. 4, 01.04.2012, p. 1101-1107.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Pyruvate protects the brain against ischemia-reperfusion injury by activating the erythropoietin signaling pathway

AU - Ryou, Myoung Gwi

AU - Liu, Ran

AU - Ren, Ming

AU - Sun, Jie

AU - Mallet, Robert T.

AU - Yang, Shao Hua

PY - 2012/4/1

Y1 - 2012/4/1

N2 - Background and Purpose-Pyruvate is known to be cytoprotective through antioxidant and anti-inflammatory mechanisms. We tested the hypothesis that pyruvate protects the brain against ischemia-reperfusion injury by inducing endogenous erythropoietin (EPO) expression. Methods-Pyruvate's protective effect was evaluated in C6 glioma cells and HT22 neuronal cells subjected to transient oxygen glucose deprivation. Cell viability (calcein AM assay) and expression of hypoxia-inducible factor-1α, EPO, Akt and Erk (immunoblot), and EPO receptor (reverse transcription-polymerase chain reaction) were analyzed. Transient focal cerebral ischemia in rats was induced by 2 hours middle cerebral artery occlusion followed by 24 hours reperfusion. Pyruvate or saline was infused from 60 minutes occlusion until 30 minutes reperfusion. Lesion volume and DNA fragmentation were assessed by 2,3,5-triphenyltetrazolium staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay, respectively. Immunoblots were conducted to determine cerebral EPO contents. Results-Pyruvate increased cell viability, hypoxia-inducible factor-1α, EPO, and Akt phosphorylation. Small interfering RNA suppression of hypoxia-inducible factor-1α and EPO abolished pyruvate-induced cytoprotection. In the rat stroke model, pyruvate reduced lesion volume by 84% and DNA fragmentation by 77% versus controls; increased EPO content paralleled these cerebroprotective actions of pyruvate. Conclusions-Pyruvate activation of the hypoxia-inducible factor-1α-EPO signaling cascade in neurons and glia could protect the brain from ischemia-reperfusion injury.

AB - Background and Purpose-Pyruvate is known to be cytoprotective through antioxidant and anti-inflammatory mechanisms. We tested the hypothesis that pyruvate protects the brain against ischemia-reperfusion injury by inducing endogenous erythropoietin (EPO) expression. Methods-Pyruvate's protective effect was evaluated in C6 glioma cells and HT22 neuronal cells subjected to transient oxygen glucose deprivation. Cell viability (calcein AM assay) and expression of hypoxia-inducible factor-1α, EPO, Akt and Erk (immunoblot), and EPO receptor (reverse transcription-polymerase chain reaction) were analyzed. Transient focal cerebral ischemia in rats was induced by 2 hours middle cerebral artery occlusion followed by 24 hours reperfusion. Pyruvate or saline was infused from 60 minutes occlusion until 30 minutes reperfusion. Lesion volume and DNA fragmentation were assessed by 2,3,5-triphenyltetrazolium staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay, respectively. Immunoblots were conducted to determine cerebral EPO contents. Results-Pyruvate increased cell viability, hypoxia-inducible factor-1α, EPO, and Akt phosphorylation. Small interfering RNA suppression of hypoxia-inducible factor-1α and EPO abolished pyruvate-induced cytoprotection. In the rat stroke model, pyruvate reduced lesion volume by 84% and DNA fragmentation by 77% versus controls; increased EPO content paralleled these cerebroprotective actions of pyruvate. Conclusions-Pyruvate activation of the hypoxia-inducible factor-1α-EPO signaling cascade in neurons and glia could protect the brain from ischemia-reperfusion injury.

KW - brain recovery

KW - erythropoietin

KW - focal ischemia

KW - hypoxia-inducible factor

KW - pyruvate

KW - stroke care

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U2 - 10.1161/STROKEAHA.111.620088

DO - 10.1161/STROKEAHA.111.620088

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