TY - JOUR
T1 - Pre-B-cell-colony-enhancing factor is critically involved in thrombin-induced lung endothelial cell barrier dysregulation
AU - Ye, Shui Q.
AU - Zhang, Li Q.
AU - Adyshev, Djanybek
AU - Usatyuk, Peter V.
AU - Garcia, Alexander N.
AU - Lavoie, Tera L.
AU - Verin, Alexander D.
AU - Natarajan, Viswanathan
AU - Garcia, Joe G.N.
N1 - Funding Information:
This work was supported by National Heart, Lung, and Blood Institute Grants (SCCOR-1P50HL 073994).
PY - 2005/11
Y1 - 2005/11
N2 - Prior genomic and genetic studies identified pre-B-cell colony-enhancing factor (PBEF) as a novel candidate gene and biomarker in acute lung injury (ALI). As increased vascular permeability is a cardinal feature of ALI, we assessed the role of PBEF in in vitro vascular barrier regulation using confluent human pulmonary artery endothelial cell (HPAEC) monolayers. Reductions in PBEF protein expression (>70%) by siRNA significantly attenuated EC barrier dysfunction induced by the potent edemagenic agent, thrombin, reflected by reductions in transendothelial electric resistance (TER, ∼60% reduction). Furthermore, PBEF siRNA blunted thrombin-mediated increases in Ca2+ entry, polymerized actin formation, and myosin light chain phosphorylation, events critical to the thrombin-mediated permeability response. Finally, PBEF siRNA also significantly inhibited thrombin-stimulated increase of IL-8 secretion in HPAEC, a chemokine known to induce actin fiber formation and intercellular gap formation of endothelial cells. Taken together, these studies demonstrate that PBEF may be required for complete expression of the thrombin-induced inflammatory response and reveal potentially novel role for PBEF in the regulation of EC Ca2+-dependent cytoskeletal rearrangement and endothelial barrier dysfunction. Ongoing studies will continue to address the molecular mechanisms by which PBEF contributes to ALI susceptibility.
AB - Prior genomic and genetic studies identified pre-B-cell colony-enhancing factor (PBEF) as a novel candidate gene and biomarker in acute lung injury (ALI). As increased vascular permeability is a cardinal feature of ALI, we assessed the role of PBEF in in vitro vascular barrier regulation using confluent human pulmonary artery endothelial cell (HPAEC) monolayers. Reductions in PBEF protein expression (>70%) by siRNA significantly attenuated EC barrier dysfunction induced by the potent edemagenic agent, thrombin, reflected by reductions in transendothelial electric resistance (TER, ∼60% reduction). Furthermore, PBEF siRNA blunted thrombin-mediated increases in Ca2+ entry, polymerized actin formation, and myosin light chain phosphorylation, events critical to the thrombin-mediated permeability response. Finally, PBEF siRNA also significantly inhibited thrombin-stimulated increase of IL-8 secretion in HPAEC, a chemokine known to induce actin fiber formation and intercellular gap formation of endothelial cells. Taken together, these studies demonstrate that PBEF may be required for complete expression of the thrombin-induced inflammatory response and reveal potentially novel role for PBEF in the regulation of EC Ca2+-dependent cytoskeletal rearrangement and endothelial barrier dysfunction. Ongoing studies will continue to address the molecular mechanisms by which PBEF contributes to ALI susceptibility.
KW - Acute lung injury
KW - Cytoskeleton
KW - Pulmonary permeability
KW - siRNA
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U2 - 10.1016/j.mvr.2005.08.003
DO - 10.1016/j.mvr.2005.08.003
M3 - Article
C2 - 16188281
AN - SCOPUS:27744546092
SN - 0026-2862
VL - 70
SP - 142
EP - 151
JO - Microvascular Research
JF - Microvascular Research
IS - 3
ER -