Hypoxia-inducible factor and nuclear factor kappa-B activation in blood-brain barrier endothelium under hypoxic/reoxygenation stress

Ken A. Witt, Karen S. Mark, Jason Huber, Thomas P. Davis

Research output: Contribution to journalArticle

43 Scopus citations

Abstract

This investigation focuses on transcription factor involvement in blood-brain barrier (BBB) endothelial cell-induced alterations under conditions of hypoxia and post-hypoxia/reoxygenation (H/R), using established in vivo/ex vivo and in vitro BBB models. Protein/DNA array analyses revealed a correlation in key transcription factor activation during hypoxia and H/R, including NFκB and hypoxia-inducible factor (HIF)1. Electrophoretic mobility shift assays confirmed NFκB and HIF1 binding activity ex vivo and in vitro, under conditions of hypoxia and H/R. Hypoxia- and H/R-treated BBB endothelium showed increased HIF1α protein expression in both cytoplasmic and nuclear fractions, in ex vivo and in vitro models. Co-immunoprecipitation of HIF1α and HIF1β was shown in the nuclear fraction under conditions of hypoxia and H/R in both models. Hypoxia- and H/R-treated BBB endothelium showed increased expression of NFκB-p65 protein in both cytoplasmic and nuclear fractions. Co-immunoprecipitation of NFκB-p65 with NFκB-p50 was shown in the nuclear fraction under conditions of hypoxia and H/R in the ex vivo model, and after H/R in the in vitro model. These data offer novel avenues in which to alter and/or investigate BBB activity across model systems and to further our understanding of upstream regulators during hypoxia and H/R.

Original languageEnglish (US)
Pages (from-to)203-214
Number of pages12
JournalJournal of neurochemistry
Volume92
Issue number1
DOIs
StatePublished - Jan 1 2005

Keywords

  • Blood-brain barrier
  • Hypoxia
  • Hypoxia-inducible factor
  • Nuclear factor kappa-B
  • Reoxygenation

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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