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Nrf2 redox signalling in brain endothelial cells adapted to physiological O2: an in vitro model for ischaemia-reperfusion injury in stroke

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Sulforaphane (SFN) has been shown to protect against damage sustained following transient ischaemic stroke via activation of the redox-sensitive transcription factor Nrf2. To further understand the protective effects of SFN… Click to show full abstract

Sulforaphane (SFN) has been shown to protect against damage sustained following transient ischaemic stroke via activation of the redox-sensitive transcription factor Nrf2. To further understand the protective effects of SFN on the cerebral endothelium, mouse brain endothelial cell line bEnd.3 was subjected to oxygen-glucose deprivation (OGD), an in vitro model of cerebral stroke. Pre-treatment of bEnd.3 cells with SFN (2.5 µM, 12 h) afforded increased protection against OGD induced cell death. Our lab recently reported that long-term culture under physiological oxygen, 5% O2, results in a phenotype markedly different to cells cultured under ambient air, 18% O2, shown by downregulation of selected Nrf2 target genes by Nrf2 activator. In the present study, the bEnd.3 cell phenotype was determined following long-term (5 days) culture under 5% or 18% O2. Under 5% O2, the intracellular O2 content of bEnd.3 cells was 3.6% O2, and a reduced rate of cell proliferation was observed. Furthermore, bEnd.3 cells cultured under 5% O2 exhibited greater sensitivity to OGD-induced cell death. The present study highlights the importance of conducting cell culture studies under physiological O2 levels.

Keywords: vitro model; redox; brain endothelial; bend cells; cell

Journal Title: Free Radical Biology and Medicine
Year Published: 2018

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