Fluid flow regulates E-selectin protein levels in human endothelial cells by inhibiting translation

Larry W. Kraiss, Neal M. Alto, Dan A. Dixon, Thomas M. McIntyre, Andrew S. Weyrich, Guy A. Zimmerman

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

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-α [TNF-α], interleukin-1β, oncostatin M, or phorbol ester) in the presence or absence of fluid flow (peak shear stress, ∼12 dynes/cm2) 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-α 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-α, 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.

Original languageEnglish (US)
Pages (from-to)161-168
Number of pages8
JournalJournal of Vascular Surgery
Volume37
Issue number1
DOIs
StatePublished - Jan 1 2003

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E-Selectin
Endothelial Cells
Proteins
Tumor Necrosis Factor-alpha
Human Umbilical Vein Endothelial Cells
Messenger RNA
Oncostatin M
Polyribosomes
Phorbol Esters
Ribonucleases
Interleukin-1
Immunoblotting
Enzyme-Linked Immunosorbent Assay

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Surgery

Cite this

Fluid flow regulates E-selectin protein levels in human endothelial cells by inhibiting translation. / Kraiss, Larry W.; Alto, Neal M.; Dixon, Dan A.; McIntyre, Thomas M.; Weyrich, Andrew S.; Zimmerman, Guy A.

In: Journal of Vascular Surgery, Vol. 37, No. 1, 01.01.2003, p. 161-168.

Research output: Contribution to journalArticle

Kraiss, Larry W. ; Alto, Neal M. ; Dixon, Dan A. ; McIntyre, Thomas M. ; Weyrich, Andrew S. ; Zimmerman, Guy A. / Fluid flow regulates E-selectin protein levels in human endothelial cells by inhibiting translation. In: Journal of Vascular Surgery. 2003 ; Vol. 37, No. 1. pp. 161-168.
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