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Am J Physiol Heart Circ Physiol 279: H2815-H2823, 2000;
0363-6135/00 $5.00
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Vol. 279, Issue 6, H2815-H2823, December 2000

TNF-alpha increases entry of macromolecules into luminal endothelial cell glycocalyx

Charmaine B. S. Henry and Brian R. Duling

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-alpha (TNF-alpha ) 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-alpha . 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-alpha 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-alpha could be observed as early as 20 min after the onset of topical application. TNF-alpha 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-alpha application, and we used the selectin antagonist fucoidan to attenuate leukocyte adhesion during TNF-alpha stimulation. This did not inhibit the TNF-alpha -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-alpha


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