OBJECTIVE: The purpose of this study was to determine the mechanism with which fluid flow inhibits endothelial E-selectin expression. METHODS: Cultured human umbilical vein endothelial cells were stimulated with inflammatory agonists (tumor necrosis factor-alpha [TNF-alpha], interleukin-1beta, oncostatin M, or phorbol ester) in the presence or absence of fluid flow (peak shear stress, approximately 12 dynes/cm(2)) imposed with an orbital shaker. E-selectin expression was assessed with ribonuclease protection assay, immunoblotting, enzyme-linked immunosorbent assay, or metabolic labeling as appropriate. RESULTS: All agonists caused human umbilical vein endothelial cells to express E-selectin protein. Fluid flow inhibited E-selectin protein levels by about 50% in response to TNF-alpha but had no effect on total E-selectin messenger RNA (mRNA) expression. Flow inhibited E-selectin protein production even after initiation of E-selectin transcription. Flow did not cause E-selectin to be shed from the cell surface nor was E-selectin degradation accelerated. Although fluid flow did not reduce total cellular E-selectin mRNA levels in response to TNF-alpha, the amount of E-selectin mRNA present in the actively translated polysome fraction was markedly attenuated. CONCLUSION: These findings indicate that E-selectin expression is subject to translational and transcriptional control. Fluid mechanical forces can regulate endothelial phenotype by targeting translational control points.
OBJECTIVE: The purpose of this study was to determine the mechanism with which fluid flow inhibits endothelial E-selectin expression. METHODS: Cultured human umbilical vein endothelial cells were stimulated with inflammatory agonists (tumor necrosis factor-alpha [TNF-alpha], interleukin-1beta, oncostatin M, or phorbol ester) in the presence or absence of fluid flow (peak shear stress, approximately 12 dynes/cm(2)) imposed with an orbital shaker. E-selectin expression was assessed with ribonuclease protection assay, immunoblotting, enzyme-linked immunosorbent assay, or metabolic labeling as appropriate. RESULTS: All agonists caused human umbilical vein endothelial cells to express E-selectin protein. Fluid flow inhibited E-selectin protein levels by about 50% in response to TNF-alpha but had no effect on total E-selectin messenger RNA (mRNA) expression. Flow inhibited E-selectin protein production even after initiation of E-selectin transcription. Flow did not cause E-selectin to be shed from the cell surface nor was E-selectin degradation accelerated. Although fluid flow did not reduce total cellular E-selectin mRNA levels in response to TNF-alpha, the amount of E-selectin mRNA present in the actively translated polysome fraction was markedly attenuated. CONCLUSION: These findings indicate that E-selectin expression is subject to translational and transcriptional control. Fluid mechanical forces can regulate endothelial phenotype by targeting translational control points.
Authors: Peter B Brant-Zawadzki; Douglas I Schmid; Huimao Jiang; Andrew S Weyrich; Guy A Zimmerman; Larry W Kraiss Journal: J Vasc Surg Date: 2007-06 Impact factor: 4.268
Authors: Douglas I Schmid; Hansjörg Schwertz; Huimiao Jiang; Robert A Campbell; Andrew S Weyrich; Thomas M McIntyre; Guy A Zimmerman; Larry W Kraiss Journal: J Cell Biochem Date: 2013-07 Impact factor: 4.429
Authors: Huimiao Jiang; Hansjörg Schwertz; Douglas I Schmid; Brandt B Jones; John Kriesel; Mark L Martinez; Andrew S Weyrich; Guy A Zimmerman; Larry W Kraiss Journal: Arterioscler Thromb Vasc Biol Date: 2012-02-09 Impact factor: 8.311