NF- kappa B independent suppression of endothelial vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 gene expression by inhibition of flavin binding proteins and superoxide production

J Mol Cell Cardiol. 2000 Aug;32(8):1499-508. doi: 10.1006/jmcc.2000.1183.


Oxidation-reduction (redox) coupled mechanisms play an important role in the regulation of cell surface adhesion molecule expression. In endothelial cells membrane-bound NADH/NADPH oxidase is a significant source of intracellular superoxide (O(2)(-)) production. We explored the role of flavin containing proteins such as NADH/NADPH oxidase in the induction of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) gene expression in human aortic endothelial cells (HAECs) and human dermal microvascular endothelial cells (HMECs). Treatment of HAECs by tumor necrosis factor- alpha (TNF- alpha, 100 U/ml) for 1 h induced a 31% increase in O(2)(-)production within 5 min as determined by lucigenin chemiluminescence analysis of whole cells (n=4, P<0.05). Pretreatment with the NADH/NADPH oxidase inhibitor diphenylene iodonium (DPI, 40 microm) for 1 h inhibited O(2)(-)production. DPI also inhibited TNF and LPS-induced VCAM-1 and ICAM-1 cell surface expression and TNF- alpha, LPS, or IL-1 beta induced VCAM-1 and ICAM-1 mRNA accumulation. However, DPI did not inhibit TNF- alpha -induced activation of nuclear NF- kappa B-like binding activity in HAECs and HMECs. Furthermore, DPI did not inhibit TNF- alpha induced transactivation of NF- kappa B-driven VCAM-1 and HIV-LTR promoter gene constructs in transiently transfected HMECs. These data suggest that flavin binding proteins such as NADH/NADPH oxidase can regulate VCAM-1 gene expression independent of NF- kappa B. Furthermore, intracellular O(2)(-)generation is not necessary for NF- kappa B activation or for transactivation of NF- kappa B driven promoters.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aorta / metabolism
  • Blotting, Northern
  • Cell Adhesion
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Endothelium, Vascular / cytology
  • Enzyme Inhibitors / pharmacology
  • Enzyme-Linked Immunosorbent Assay
  • Flavins / metabolism*
  • Gene Expression / drug effects*
  • HIV Long Terminal Repeat / genetics
  • Humans
  • Intercellular Adhesion Molecule-1 / metabolism*
  • NF-kappa B / metabolism*
  • Onium Compounds / pharmacology
  • Oxidation-Reduction
  • Promoter Regions, Genetic
  • Protein Binding
  • RNA, Messenger / metabolism
  • Superoxides / antagonists & inhibitors*
  • Time Factors
  • Transcriptional Activation
  • Transfection
  • Tumor Necrosis Factor-alpha / metabolism
  • Vascular Cell Adhesion Molecule-1 / metabolism*


  • Enzyme Inhibitors
  • Flavins
  • NF-kappa B
  • Onium Compounds
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha
  • Vascular Cell Adhesion Molecule-1
  • Superoxides
  • Intercellular Adhesion Molecule-1
  • diphenyleneiodonium