Hypoxia-Induced Hyperpermeability in Brain Microvessel Endothelial Cells Involves VEGF-Mediated Changes in the Expression of Zonula Occludens-1

Fischer, Silvia; Wobben, Maria; Marti, Hugo H.; Renz, D.; Schäper, Wolfgang

doi:10.1006/mvre.2001.2367

Cited by 267 publications

(214 citation statements)

References 41 publications

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“…Apoptotic cell death of cultured cerebral endothelium in response to hypoxia or simulated ischemia is not a novel finding (Beetsch et al, 1998;Bresgen et al, 2003;Fischer et al, 2002), but to our knowledge, this is the first report mechanistically linking the demise of these cells to PARP-1 activation. Although PARP-1 activation may also be triggered by phospholipase C (Homburg et al, 2000) and other signals (Koedel et al, 2002), if DNA strand breaks are the primary activator of the enzyme (de Murcia and de Murcia, 1994), then HBMECs must be vulnerable to significant chromatin damage by HX/RO stress.…”

Section: Discussionmentioning

confidence: 79%

Cerebral Endothelial Cell Apoptosis after Ischemia—Reperfusion: Role of PARP Activation and AIF Translocation

Zhang

Park

et al. 2005

J Cereb Blood Flow Metab

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Cerebral ischemia-reperfusion leads to vascular dysfunction characterized by endothelial cell injury or death. In the present study, we used an in vitro model to elucidate mechanisms of human brain microvascular endothelial cell (HBMEC) injury after episodic ischemia-reperfusion. Near-confluent HBMEC cultures were exposed to intermittent hypoxia-reoxygenation (HX/RO) and, at different recovery time points, cell viability was assessed by the MTT assay, apoptotic death by fluorescence microscopy of terminal deoxynucleotidyl transferase-mediated 2 0 -deoxyuridine 5 0 -triphosphatebiotin nick end labeling (TUNEL)-positive cells, and nuclear translocation of apoptosis-inducing factor (AIF) and cleavage of poly(ADP-ribose) polymerase-1 (PARP-1) by immunoblotting of subcellular fractions. Reductions in HBMEC viability were proportional to the number of HX/RO cycles, and not the total duration of hypoxia. Using four cycles of 1-h HX with 1 h of intervening normoxic RO, cell viability was reduced 30% to 40% between 12 and 48 h. Treatment with the PARP-1 inhibitors 3-aminobenzamide or 4-amino-1,8-naphthalimide during the insult improved HBMEC viability at 24 h after insult, and resulted in dose-dependent reductions in TUNEL-positivity at 16 h after insult, but not if these treatments were delayed by 4 h. HX/RO-induced increases in nuclear AIF translocation, as well as PARP-1 cleavage, were also reduced dose-dependently at 4 h after insult by the inhibitors. The caspase inhibitor z-VAD-fmk blocked PARP-1 cleavage, but did not affect AIF translocation and was only modestly cytoprotective. These findings indicate that PARP-1 activation and a PARP-1-dependent, caspase-independent, nuclear translocation of AIF contribute to apoptotic cerebral endothelial cell death after ischemia-reperfusion, underscoring the potential for ischemic microvascular protection by inhibiting PARP activation or preventing AIF translocation.

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Section: Discussionmentioning

confidence: 79%

Cerebral Endothelial Cell Apoptosis after Ischemia—Reperfusion: Role of PARP Activation and AIF Translocation

Zhang

Park

et al. 2005

J Cereb Blood Flow Metab

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“…Nonetheless, the immunoprecipitation experiments demonstrate that despite the low expression, CLDN1 directly binds to ZO‐1 in HBMECs, and this association becomes more efficient after the miR‐155 downregulation. ZO‐1 levels and localization are influenced by its phosphorylation 8, 34. Therefore, we compared the ZO‐1 phosphorylation state in anti–ZO‐1 immunoprecipitates from OGD/I and OGD/IC cells.…”

Section: Resultsmentioning

confidence: 99%

“…Stroke results in the breakdown of the BBB, with subsequent damage to cerebral vasculature and nonvascular brain elements, including neurons, astrocytes, and microglia. Hypoxia‐induced vascular damage is associated with disruption of paracellular cell junctions, with selective targeting of brain microvascular endothelial TJ complexes 6, 7, 8…”

Section: Introductionmentioning

confidence: 99%

Inhibition of MicroRNA‐155 Supports Endothelial Tight Junction Integrity Following Oxygen‐Glucose Deprivation

Peña-Philippides

Gardiner

Caballero-Garrido

et al. 2018

JAHA

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BackgroundBrain microvascular endothelial cells form a highly selective blood brain barrier regulated by the endothelial tight junctions. Cerebral ischemia selectively targets tight junction protein complexes, which leads to significant damage to cerebral microvasculature. Short noncoding molecules called microRNAs are implicated in the regulation of various pathological states, including endothelial barrier dysfunction. In the present study, we investigated the influence of microRNA‐155 (miR‐155) on the barrier characteristics of human primary brain microvascular endothelial cells (HBMECs).Methods and ResultsOxygen‐glucose deprivation was used as an in vitro model of ischemic stroke. HBMECs were subjected to 3 hours of oxygen‐glucose deprivation, followed by transfections with miR‐155 inhibitor, mimic, or appropriate control oligonucleotides. Intact normoxia control HBMECs and 4 oxygen‐glucose deprivation–treated groups of cells transfected with appropriate nucleotide were subjected to endothelial monolayer electrical resistance and permeability assays, cell viability assay, assessment of NO and human cytokine/chemokine release, immunofluorescence microscopy, Western blot, and polymerase chain reaction analyses. Assessment of endothelial resistance and permeability demonstrated that miR‐155 inhibition improved HBMECs monolayer integrity. In addition, miR‐155 inhibition significantly increased the levels of major tight junction proteins claudin‐1 and zonula occludens protein‐1, while its overexpression reduced these levels. Immunoprecipitation and colocalization analyses detected that miR‐155 inhibition supported the association between zonula occludens protein‐1 and claudin‐1 and their stabilization at the HBMEC membrane. Luciferase reporter assay verified that claudin‐1 is directly targeted by miR‐155.ConclusionsBased on these results, we conclude that miR‐155 inhibition–induced strengthening of endothelial tight junctions after oxygen‐glucose deprivation is mediated via its direct target protein claudin‐1.

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“…BBB VEGF was initially described as a vascular permeability factor, and one of the main effects of VEGF intracortical administration is the increase in BBB permeability in adults (Dobrogowska et al, 1998;Fischer et al, 2002;Krum et al, 2002). Previous studies have posited EBA as a specific marker for the BBB in rats, linking its expression with the functionality of the blood-brain barrier (Lin et al, 2001;Norsted et al, 2008) since a decrease in EBA positivity has been reported in pathological conditions where the administration of an anti-EBA antibody resulted in increased permeability of brain microvessels (Ghabriel et al, 2000;Ghabriel et al, 2002).…”

Section: 1mentioning

confidence: 99%

“…Outside the brain, the blockade of VEGF by polyclonal antibodies impedes the fracture healing process (Chu et al, 2009). Authors report that VEGF plays an important role as an endogenous cytoprotective mechanism and that VEGF induces both BBB permeability and inflammation on a dose-dependent basis (Dobrogowska et al, 1998;Fischer et al, 2002;Croll et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Effect of intracortical vascular endothelial growth factor infusion and blockade during the critical period in the rat visual cortex

et al. 2012

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Hypoxia-Induced Hyperpermeability in Brain Microvessel Endothelial Cells Involves VEGF-Mediated Changes in the Expression of Zonula Occludens-1

Cited by 267 publications

References 41 publications

Cerebral Endothelial Cell Apoptosis after Ischemia—Reperfusion: Role of PARP Activation and AIF Translocation

Cerebral Endothelial Cell Apoptosis after Ischemia—Reperfusion: Role of PARP Activation and AIF Translocation

Inhibition of MicroRNA‐155 Supports Endothelial Tight Junction Integrity Following Oxygen‐Glucose Deprivation

Effect of intracortical vascular endothelial growth factor infusion and blockade during the critical period in the rat visual cortex

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