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Vol. 297, Issue 3, 1051-1058, June 2001
Induces Cyclooxygenase-2 Expression and
Prostaglandin Release in Brain Microvessel Endothelial Cells
Department of Pharmaceutical Science, College of Pharmacy,
University of Nebraska Medical Center, Omaha, Nebraska
Primary cultured bovine brain microvessel endothelial cells (BBMECs),
were used as an in vitro model of the blood-brain barrier to examine
the involvement of eicosanoids in the permeability and cytoskeletal
structural changes observed following exposure to tumor necrosis
factor-
(TNF-). Compared with control monolayers, BBMECs exposed
to TNF- formed actin filament tangles and extracellular gaps with a
resultant increase in permeability. Both the permeability and
cytoskeletal changes observed with TNF- were significantly reduced
following pretreatment with NS-398 or indomethacin, inhibitors of
cyclooxygenase (COX). Western blot analysis showed that TNF- had no
apparent effect on the expression of COX-1, but did induce the
expression of COX-2 in the BBMECs. The induction of COX-2 expression
occurred within the same time frame (2-4 h following TNF- exposure)
as the permeability increases observed with the cytokine. Consistent
with the increased expression of COX-2, BBMEC monolayers exposed to
TNF- had significantly greater secretion and release of
prostaglandin E2 (PGE2) and prostaglandin
F2
(PGF2). Furthermore, BBMEC monolayers
treated with PGE2 or PGF2 showed significant
increases in permeability and cytoskeletal structural changes when
compared with control monolayers. Together, these results suggest that
the TNF--induced permeability and cytoskeletal structural effects
are due, in part, to an induction of the COX-2 system and increased
release of prostaglandins in the cerebral microvasculature.
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