Background
Gram-negative bacterial lipopolysaccharide (LPS) leads to the pathologic increase
of vascular leakage under septic conditions. However, the mechanisms behind LPS-induced
vascular hyperpermeability remain incompletely understood. In this study, we tested
hypothesis that guanine nucleotide exchange factor-H1 (GEF-H1) signaling might be
a key pathway involved in endothelial cells (ECs) barrier dysfunction.
Methods
The roles of GEF-H1 signaling pathway in LPS-induced ECs barrier dysfunction were
accessed by Evans blue dye-labeled albumin (EB-albumin) leak across the human umbilical
vein EC (HUVEC) monolayers and Western blot assays. Furthermore, the effect of GEF-H1
signaling on LPS-induced alteration of cytoskeletal proteins and disruption of cell–cell
junctions were analyzed by immunofluorescent analysis and Western blot assays, respectively.
Results
We found that LPS could rapidly activated GEF-H1/RhoA/Rho-associated protein kinase
(ROCK) signaling pathway in ECs. The LPS-mediated increase in EB-albumin flux across
human HUVECs monolayers could be prevented by GEF-H1 depletion or ROCK inactivation.
ECs permeability is controlled by actin filaments and cell–cell contact protein complexes.
Actin stress fiber formation and/or cell–cell contact proteins loss cause vascular
barrier disruption. Here, GEF-H1 knockdown or ROCK inactivation both not only significantly
inhibited LPS-induced actin stress fiber formation, phosphorylation of myosin light
chain, and myosin-associated phosphatase type 1, but also suppressed LPS-induced loss
of occludin, claudin-1, and vascular endothelial (VE)-cadherin in ECs, which suggested
that LPS-induced stress fiber formation and cell–cell junctions disruption were closely
associated with GEF-H1/RhoA/ROCK signaling activation.
Conclusion
Our findings indicate that GEF-H1/RhoA/ROCK pathway in ECs plays an important role
in LPS-mediated alteration of cell morphology and disruption of cell–cell junctions,
consequently regulate LPS-induced vascular permeability dysfunction.
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Article info
Publication history
Published online: July 15, 2013
Accepted:
April 3,
2013
Footnotes
Z.Z. and F.G. contributed equally to this work.
Supported by the National Natural Science Foundation of China (No.81071553 and No.81272094).
Identification
Copyright
© 2013 Mosby, Inc. Published by Elsevier Inc. All rights reserved.
