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Molecular and Cellular Biochemistry

, Volume 367, Issue 1–2, pp 31–42 | Cite as

Post-transcriptional regulation of VEGF-A mRNA levels by mitogen-activated protein kinases (MAPKs) during metabolic stress associated with ischaemia/reperfusion

  • Bryan W. Miller
  • Joanna M. Hay
  • Sally A. Prigent
  • Martin DickensEmail author
Article

Abstract

Angiogenesis is a well-characterised response to the metabolic stresses that occur during ischaemia/reperfusion, but the signalling pathways that regulate it are poorly understood. We tested whether activation of mitogen-activated protein kinases (MAPKs) was involved in regulating the expression of pro-angiogenic growth factors by the metabolic stresses associated with ischaemia/reperfusion in H9c2 rat cardiomyoblasts. Metabolic stress had no effect on vascular endothelial growth factor (VEGF) mRNA levels, but recovery after metabolic inhibition led to a strong induction of VEGF-A mRNA (3.8 ± 0.5-fold at 4 h), a modest rise in VEGF-C mRNA levels (1.7 ± 0.3-fold at 4 h), with no effect on VEGF-B or -D. A VEGF-A promoter reporter construct was unresponsive to metabolic inhibition/recovery and increases in VEGF-A mRNA were not blocked by the transcription inhibitor actinomycin D suggesting that increases in VEGF mRNA were due to enhanced VEGF-A mRNA stability. In addition, studies using reporter constructs demonstrated that regions within the 5′ untranslated region (UTR) contributed to enhanced mRNA stability following recovery from metabolic stress. Increases in VEGF-A mRNA were abolished by inhibition of extracellular signal-regulated kinase or c-jun N-terminal kinase MAPKs, suggesting that these kinases may promote angiogenesis in response to metabolic stress during ischaemia/reperfusion by increasing VEGF-A message stability.

Keywords

Vascular endothelial growth factor Angiogenesis Metabolic stress Mitogen-activated protein kinase 

Abbreviations

ATF2

Activating transcription factor 2

ATP

Adenosine 5′ triphosphate

CAT

Chloramphenicol acetyl transferase

dCTP

Deoxy cytosine 5′ triphosphate

ERK

Extracellular signal-regulated kinase

JNK

c-jun N-terminal kinase

MAPK

Mitogen-activated protein kinase

MKK6

MAPK kinase 6

PMA

Phorbol myristoyl acetate

RT-PCR

Reverse transcription polymerase chain reaction

SEM

Standard error of the mean

VEGF

Vascular endothelial growth factor

Notes

Acknowledgments

This work was supported, in part, by a grant from the British Heart Foundation (PG#98111) and studentships from the Biotechnology and Biological Sciences Research Council (#08170) and Medical Research Council (#G78/6321). We would like to thank Dr. Ben Zion-Levy for providing us with the pV3.4-CAT construct.

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Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Bryan W. Miller
    • 1
  • Joanna M. Hay
    • 1
  • Sally A. Prigent
    • 1
  • Martin Dickens
    • 1
    Email author
  1. 1.Department of BiochemistryUniversity of LeicesterLeicesterUK

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