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Reversible oxidant-induced increases in albumin transfer across cultured
endothelium: alterations in cell shape and calcium homeostasis
DM Shasby, SE Lind, SS Shasby, JC Goldsmith and GW Hunninghake
To determine whether reactive oxygen molecules could directly and
reversibly increase the transfer of albumin across an endothelial barrier,
we measured albumin transfer across monolayers of endothelium cultured on
micropore filters before and after exposure to xanthine and xanthine
oxidase. Xanthine and xanthine oxidase increased endothelial albumin
transfer in a dose-dependent fashion. Parallel phase contrast and
fluorescence microscopy demonstrated retraction of adjacent cells from one
another and disruption of the actin filaments. The oxidant- induced
increases in albumin transfer and changes in cell shape were reversed by
removing xanthine oxidase and then incubating the monolayers for 3 1/2
hours in tissue culture media enriched with fetal bovine serum. However,
incubation in tissue culture media without serum resulted in progressive
injury and cell death. Hence, the brief exposure to oxidants initiated a
progressive injury process that was reversed by incubation in serum.
Because intracellular and extracellular calcium are important determinants
of cell shape, and because some oxidized membrane lipids act as calcium
ionophores, we asked whether oxidants altered endothelial calcium
homeostasis. Xanthine-xanthine oxidase increased release of 45Ca++ from
preloaded cells. The calcium antagonist lanthanum chloride prevented
xanthine- xanthine oxidase increases in endothelial albumin transfer and
prevented the changes in cell shape; chelation of extracellular calcium
inhibited lysis of endothelium by xanthine-xanthine oxidase; and the
calcium ionophore A23187 increased endothelial albumin transfer and
mimicked the oxidant-induced changes in cell shape. Lanthanum chloride
inhibited these effects of A23187. These data suggest that oxygen radicals
can reversibly increase endothelial permeability to macromolecules, that
this is associated with reversible changes in endothelial cell shape and
actin filaments, and that the changes in cell shape are related to
oxidant-induced changes in endothelial calcium homeostasis.
Volume 65,
Issue 3,
pp. 605-614,
03/01/1985
Copyright © 1985 by The American Society of Hematology
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