Inhibition of VCAM-1 expression in endothelial cells by CORM-3: the role of the ubiquitin-proteasome system, p38, and mitochondrial respiration

Free Radic Biol Med. 2012 Feb 15;52(4):794-802. doi: 10.1016/j.freeradbiomed.2011.11.035. Epub 2011 Dec 21.

Abstract

Carbon monoxide (CO) abrogates TNF-α-mediated inflammatory responses in endothelial cells, yet the underlying mechanism thereof is still elusive. We have previously shown that the anti-inflammatory effect of CO-releasing molecule-3 (CORM-3) is not completely mediated via deactivation of the NF-κB pathway. In this study, we sought to explore other potential mechanisms by which CORM-3 downregulates VCAM-1 expression on TNF-α-stimulated HUVECs. By genome-wide gene expression profiling and pathway analysis we studied the relevance of particular pathways for the anti-inflammatory effect of CORM-3. In CORM-3-stimulated HUVECs significant changes in expression were found for genes implicated in the proteasome and porphyrin pathways. Although proteasome activities were increased by CORM-3, proteasome inhibitors did not abolish the effect of CORM-3. Likewise, heme oxygenase-1 inhibitors did not abrogate the ability of CORM-3 to downregulate VCAM-1 expression. Interestingly, CORM-3 inhibited MAPK p38, and the p38 inhibitor SB203580 downregulated VCAM-1 expression. However, downregulation of VCAM-1 by CORM-3 occurred only at concentrations that partly inhibit ATP production and sodium azide and oligomycin paralleled the effect of CORM-3 in this regard. Our results indicate that CORM-3-induced downregulation of VCAM-1 is mediated via p38 inhibition and mitochondrial respiration, whereas the ubiquitin-proteasome system seems not to be involved.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Inflammatory Agents / pharmacology*
  • Carbon Monoxide / pharmacology
  • Cells, Cultured
  • Chemokines / genetics
  • Chemokines / metabolism
  • Down-Regulation
  • Enzyme Activation
  • Gene Expression / drug effects*
  • Gene Expression Profiling
  • Human Umbilical Vein Endothelial Cells / drug effects*
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Inflammation / metabolism
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism
  • MAP Kinase Signaling System*
  • Mitochondria / drug effects
  • Mitochondria / metabolism*
  • Oligonucleotide Array Sequence Analysis
  • Organometallic Compounds / pharmacology*
  • Oxygen Consumption
  • Proteasome Endopeptidase Complex / genetics
  • Proteasome Endopeptidase Complex / metabolism*
  • Proteolysis
  • Tumor Necrosis Factor-alpha / pharmacology
  • Ubiquitinated Proteins / metabolism
  • Vascular Cell Adhesion Molecule-1 / genetics
  • Vascular Cell Adhesion Molecule-1 / metabolism*

Substances

  • Anti-Inflammatory Agents
  • Chemokines
  • Intracellular Signaling Peptides and Proteins
  • Organometallic Compounds
  • Tumor Necrosis Factor-alpha
  • Ubiquitinated Proteins
  • Vascular Cell Adhesion Molecule-1
  • tricarbonylchloro(glycinato)ruthenium(II)
  • Carbon Monoxide
  • Proteasome Endopeptidase Complex