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Hypertonic mannitol loading of NF-kappaB transcription factor decoys in human brain microvascular endothelial cells blocks upregulation of ICAM-1.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Hess, David C. Howard, Edward A. Cheng, Ching-Wen Carroll, John W. Hill, William D. Hsu, Chung Y. |
| Copyright Year | 2000 |
| Abstract | BACKGROUND AND PURPOSE An acute inflammatory response exacerbates tissue injury during acute ischemic stroke. The transcription factor nuclear factor (NF)-kappaB plays a key role in endothelial cell activation and the inflammatory response. Targeted genetic disruption of NF-kappaB activation in cerebral endothelial cells may be protective in stroke. We determined whether a NF-kappaB 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)-alpha and conditions of hypoxia-reoxygenation (H/R). Mannitol was used to load phosphothiorated oligonucleotides containing 3 copies of the kappaB binding sequences (TFDs) into cultured HBMEC. An NF-kappaB TFD, a mutated NF-kappaB 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-kappaB TFD but not the mutated or scrambled TFD competed with a kappaB sequence for binding to nuclear extracts from HBMEC exposed to TNF-alpha. The NF-kappaB TFD blocked the TNF-alpha-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-kappaB decoy blocks both TNF-alpha-induced and H/R-induced ICAM-1 upregulation in HBMEC. Targeted genetic disruption of endothelial NF-kappaB activation may be of benefit in acute ischemic stroke. |
| File Format | PDF HTM / HTML |
| DOI | 10.1161/01.STR.31.5.1179 |
| Alternate Webpage(s) | http://stroke.ahajournals.org/content/strokeaha/31/5/1179.full.pdf |
| Alternate Webpage(s) | http://stroke.ahajournals.org/content/strokeaha/31/5/1179.full.pdf?download=true |
| PubMed reference number | 10797183 |
| Alternate Webpage(s) | https://doi.org/10.1161/01.STR.31.5.1179 |
| Journal | Medline |
| Volume Number | 31 |
| Issue Number | 5 |
| Journal | Stroke |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
