The MAP kinase pathway mediates transcytosis induced by TNF-alpha in an in vitro blood-brain barrier model.
Cerebral capillary endothelial cells constitute the blood-brain barrier (BBB). In these highly specialized cells, transcellular transports rarely occur, and the presence of tight junctions between them leads to a low paracellular permeability. In order to understand the functions of this barrier, an in vitro model of the BBB has been developed and consists in a co-culture of primary cerebral capillary endothelial cells and glial cells. When these endothelial cells are subjected to an inflammatory agent, such as tumor necrosis factor-alpha (TNF-alpha), in vitro BBB permeability is increased, as indicated by the increase in holotransferrin transcytosis. However, no significant change in the paracellular permeability is observed. In order to understand the molecular mechanisms that underlie these transcytosis processes, we investigated the implication of the mitogen-activated protein kinase (MAPK) signalling pathway, as TNF-alpha is known to activate this kinase family. In the present study, an increase in the activation of p42-44 MAPK is observed after TNF-alpha treatment. Holotransferrin transcytosis as well as p42-44 MAPK activation are inhibited after addition of a p42-44 MAPK pathway inhibitor (UO126) during TNF-alpha challenge. These data suggest that the MAPK pathway is involved in the transcytosis regulation in endothelial cells from an in vitro BBB model.
Miller
Florence
F
Laboratoire de Physiopathologie de la Barrière Hémato-Encéphalique, Université d'Artois EA2465, Faculté des Sciences Jean Perrin, rue Jean Souvraz, 62307 Lens cedex, France.
Fenart
Laurence
L
Landry
Valérie
V
Coisne
Caroline
C
Cecchelli
Roméo
R
Dehouck
Marie-Pierre
MP
Buée-Scherrer
Valérie
V
D000199:Actins / Q000378:metabolism; D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D001812:Blood-Brain Barrier / Q000187:drug effects* / Q000378:metabolism*; D015153:Blotting, Western / Q000379:methods; D002070:Butadienes / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D004347:Drug Interactions; D042783:Endothelial Cells / Q000187:drug effects; D004730:Endothelium, Vascular / Q000166:cytology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007150:Immunohistochemistry / Q000379:methods; D008353:Mannitol / Q000494:pharmacology; D008565:Membrane Proteins / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048290:Mitogen-Activated Protein Kinase 11 / Q000378:metabolism; D048055:Mitogen-Activated Protein Kinase 8 / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D009457:Neuroglia / Q000187:drug effects; D009570:Nitriles / Q000494:pharmacology; D062793:Occludin; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015398:Signal Transduction / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D000199:Actins; D002070:Butadienes; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D009570:Nitriles; D062793:Occludin; C568987:Ocln protein, rat; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; C113580:U 0126; D008353:Mannitol; D019950:Mitogen-Activated Protein Kinase 1; D048290:Mitogen-Activated Protein Kinase 11; D048055:Mitogen-Activated Protein Kinase 8; D020928:Mitogen-Activated Protein Kinases