Direct-contact co-culture between smooth muscle and endothelial cells inhibits TNF-alpha-mediated endothelial cell activation

Am J Physiol Heart Circ Physiol. 2010 Aug;299(2):H338-46. doi: 10.1152/ajpheart.01029.2009. Epub 2010 May 21.


We used a direct-contact endothelial cell-smooth muscle cell (EC-SMC) co-culture to examine whether quiescent SMCs regulate the EC inflammatory response to tumor necrosis factor (TNF)-alpha. ECs were cultured under static and physiological flow conditions. Compared with TNF-alpha-treated ECs in monoculture, TNF-alpha-treated ECs in co-culture had less NF-kappaB nuclear translocation; less intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin surface protein expression; no change in TNF receptor expression, but greater Kruppel-like factor 2 (KLF2) gene expression. After flow preconditioning for 24 h at 15 dyne/cm(2), and exposure of ECs to flow and TNF-alpha for 4.5 h, ECs in co-culture had less ICAM-1, VCAM-1, and E-selectin surface protein expression. Exposure to flow greatly increased KLF2 gene expression levels in both EC cultures; as a result, ECs in co-culture and monoculture had similar levels of post-flow KLF2 gene expression. The reduced levels of TNF-alpha-induced adhesion molecule expression in co-culture required the presence of quiescent SMCs; adhesion to decellularized extracellular matrix (ECM) or co-culture with fibroblasts produced only a modest reduction in EC adhesion molecule expression. Furthermore, co-culture of quiescent SMCs and ECs on the opposite side of a 10-microm-thick porous membrane did not alter the TNF-alpha-mediated ICAM-1 surface protein expression. Although the ECM produced by SMCs plays some role in reducing TNF-alpha-mediated inflammation, these results suggest that the direct contact between ECs and quiescent SMCs is required to inhibit TNF-alpha-mediated activation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Animals
  • Cell Adhesion
  • Cell Communication*
  • Cell Proliferation
  • Cellular Senescence*
  • Coculture Techniques
  • E-Selectin / metabolism
  • Endothelial Cells / immunology*
  • Endothelial Cells / metabolism
  • Extracellular Matrix / metabolism
  • Fibroblasts / metabolism
  • Humans
  • Inflammation Mediators / metabolism*
  • Intercellular Adhesion Molecule-1 / metabolism
  • Kruppel-Like Transcription Factors / metabolism
  • Mice
  • Muscle, Smooth, Vascular / cytology
  • Muscle, Smooth, Vascular / immunology*
  • Muscle, Smooth, Vascular / metabolism
  • Myocytes, Smooth Muscle / immunology*
  • Myocytes, Smooth Muscle / metabolism
  • NF-kappa B / metabolism
  • Pulsatile Flow
  • Receptors, Tumor Necrosis Factor / metabolism
  • Recombinant Proteins / metabolism
  • Stress, Mechanical
  • Time Factors
  • Tumor Necrosis Factor-alpha / metabolism*
  • Vascular Cell Adhesion Molecule-1 / metabolism


  • E-Selectin
  • Inflammation Mediators
  • KLF2 protein, human
  • Kruppel-Like Transcription Factors
  • NF-kappa B
  • Receptors, Tumor Necrosis Factor
  • Recombinant Proteins
  • Tumor Necrosis Factor-alpha
  • Vascular Cell Adhesion Molecule-1
  • Intercellular Adhesion Molecule-1