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  NO and oxyradical metabolism in new cell lines of rat brain capillary endothelial cells forming the blood–brain barrier

Blasig, I. E., Giese, H., Schroeter, M. L., Sporbert, A., Utepbergenov, D. I., Buchwalow, I. B., et al. (2001). NO and oxyradical metabolism in new cell lines of rat brain capillary endothelial cells forming the blood–brain barrier. Microvascular Research, 62(2), 114-127. doi:10.1006/mvre.2001.2318.

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 Creators:
Blasig, Ingolf E.1, Author
Giese, Helga1, Author
Schroeter, Matthias L.1, Author           
Sporbert, Anje1, Author
Utepbergenov, Darkhan I.1, Author
Buchwalow, Igor B.1, Author
Neubert, Katrin1, Author
Schönfelder, Gilbert1, Author
Freyer, Dorette1, Author
Schimke, Ingolf1, Author
Siems, Wolf-Eberhard1, Author
Paul, Martin1, Author
Haseloff, Reiner F.1, Author
Blasig, Rosel1, Author
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1External Organizations, ou_persistent22              

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Free keywords: nitric oxide; reactive oxygen species; immortalization; blood–brain barrier; brain capillary endothelial cells
 Abstract: To investigate the relevance of •NO and oxyradicals in the blood–brain barrier (BBB), differentiated and well-proliferating brain capillary endothelial cells (BCEC) are required. Therefore, rat BCEC (rBCEC) were transfected with immortalizing genes. The resulting lines exhibited endothelial characteristics (factor VIII, angiotensin-converting enzyme, high prostacyclin/thromboxane release rates) and BBB markers (γ-glutamyl transpeptidase, alkaline phosphatase). The control line rBCEC2 (mock transfected) revealed fibroblastoid morphology, less factor VIII, reduced γ-glutamyl transpeptidase, weak radical defence, low prostanoid metabolism, and limited proliferation. Lines transfected with immortalizing genes (especially rBCEC4, polyoma virus large T antigen) conserved primary properties: epitheloid morphology, subcultivation with high proliferation rate under pure culture conditions, and powerful defence against reactive oxygen species (Mn–, Cu/Zn–superoxide dismutase, catalase, glutathione peroxidase, glutathione) effectively controlling radical metabolism. Only 100 μM H2O2 overcame this defence and stimulated the formation of eicosanoids similarly as in primary cells. Some BBB markers were expressed to a lower degree; however, cocultivation with astrocytes intensified these markers (e.g., alkaline phosphatase) and paraendothelial tightness, indicating induction of BBB properties. Inducible NO synthase was induced by a cytokine plus lipopolysaccharide mixture in all lines and primary cells, resulting in •NO release. Comparing the cell lines obtained, rBCEC4 are stable immortalized and reveal the best conservation of properties from primary cells, including enzymes producing or decomposing reactive species. These cells can be subcultivated in large amounts and, hence, they are suitable to study the role of radical metabolism in the BBB and in the cerebral microvasculature.

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Language(s): eng - English
 Dates: 2000-07-282002-05-252001-09
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1006/mvre.2001.2318
PMID: 11516240
 Degree: -

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Title: Microvascular Research
  Other : Microvasc. Res.
Source Genre: Journal
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Publ. Info: Orlando, Fla. : Academic Press
Pages: - Volume / Issue: 62 (2) Sequence Number: - Start / End Page: 114 - 127 Identifier: ISSN: 0026-2862
CoNE: https://pure.mpg.de/cone/journals/resource/954922646056