Hypertonic Mannitol Loading of NF-κB Transcription Factor Decoys in Human Brain Microvascular Endothelial Cells Blocks Upregulation of ICAM-1
| Autor: | D C, Hess, E, Howard, C, Cheng, J, Carroll, W D, Hill, C Y, Hsu |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Endothelium
chemistry.chemical_compound Downregulation and upregulation Humans Medicine Transcription factor Cells Cultured Advanced and Specialized Nursing ICAM-1 Tumor Necrosis Factor-alpha business.industry NF-kappa B NF-κB Intercellular Adhesion Molecule-1 NFKB1 Cell Hypoxia Up-Regulation Endothelial stem cell medicine.anatomical_structure chemistry Reperfusion Injury Immunology Cancer research Tumor necrosis factor alpha Endothelium Vascular Neurology (clinical) Cardiology and Cardiovascular Medicine business |
| Zdroj: | Stroke. 31:1179-1186 |
| ISSN: | 1524-4628 0039-2499 |
| Popis: | Background and Purpose —An acute inflammatory response exacerbates tissue injury during acute ischemic stroke. The transcription factor nuclear factor (NF)-κB plays a key role in endothelial cell activation and the inflammatory response. Targeted genetic disruption of NF-κB activation in cerebral endothelial cells may be protective in stroke. We determined whether a NF-κB transcription factor decoy (TFD) could block intercellular adhesion molecule (ICAM)-1 upregulation, an indicator of endothelial cell activation. Methods —We modeled ischemia-reperfusion in vitro by exposing cultured human brain microvascular endothelial cells (HBMEC) to tumor necrosis factor (TNF)-α and conditions of hypoxia-reoxygenation (H/R). Mannitol was used to load phosphothiorated oligonucleotides containing 3 copies of the κB binding sequences (TFDs) into cultured HBMEC. An NF-κB TFD, a mutated NF-κB TFD, and a scrambled TFD were studied for their effect on ICAM-1 mRNA levels and surface ICAM-1 by ELISA. Results —Hyperosmolar loading with mannitol permitted rapid transfection of TFD into endothelial cell nuclei. The NF-κB TFD but not the mutated or scrambled TFD competed with a κB sequence for binding to nuclear extracts from HBMEC exposed to TNF-α. The NF-κB TFD blocked the TNF-α–induced and H/R-induced increase in ICAM-1 mRNA levels and the upregulation of surface ICAM-1. Conclusions —Mannitol delivers phosphothiorated oligonucleotides into cultured HBMEC. An NF-κB decoy blocks both TNF-α–induced and H/R-induced ICAM-1 upregulation in HBMEC. Targeted genetic disruption of endothelial NF-κB activation may be of benefit in acute ischemic stroke. |
| Databáze: | OpenAIRE |
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