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increases entry of macromolecules into
luminal endothelial cell glycocalyx
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia 22906
The endothelial luminal glycocalyx has been largely
ignored as a target in vascular pathophysiology even though it occupies a key location. As a model of the inflammatory response, we tested the
hypothesis that tumor necrosis factor-
(TNF-) can alter the
properties of the endothelial apical glycocalyx. In the intact hamster
cremaster microcirculation, fluorescein isothiocyanate (FITC)-labeled
Dextrans 70, 580, and 2,000 kDa are excluded from a region extending
from the endothelial surface almost 0.5 µm into the lumen. This
exclusion zone defines the boundaries of the glycocalyx. Red blood
cells (RBC) under normal flow conditions are excluded from a region
extending even farther into the lumen. The cremaster microcirculation
was pretreated with topical or intrascrotal applications of TNF-.
After infusion of FITC-dextran, FITC-albumin, or FITC-immunoglubulin G
(IgG) via a femoral cannula, microvessels were observed with
bright-field and fluorescence microscopy to obtain estimates of the
anatomic diameters and the widths of fluorescent tracer columns and of
the RBC columns (means ± SE). After 2 h of intrascrotal
TNF- exposure, there was a significant increase in access of
FITC-Dextrans 70 and 580 to the space bounded by the apical glycocalyx
in arterioles, capillaries, and venules, but no significant change in
access of FITC-Dextran 2,000. The effects of TNF- could be observed
as early as 20 min after the onset of topical application. TNF-
treatment also significantly increased the penetration rate of
FITC-Dextran 40, FITC-albumin, and FITC-IgG into the glycocalyx and
caused a significant increase in the intraluminal volume occupied by
flowing RBC. White blood cell adhesion increased during TNF-
application, and we used the selectin antagonist fucoidan to attenuate
leukocyte adhesion during TNF- stimulation. This did not inhibit the
TNF--mediated increase in permeation of the glycocalyx. These
results show that proinflammatory cytokines can cause disruption of the
endothelial apical glycocalyx, leading to an increased macromolecular
permeation in the absence of an increase in leukocyte recruitment.
inflammation; intravital microscopy; fluorescein
isothiocyanate-dextrans; plasma proteins; leukocytes; tumor necrosis
factor-
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