Inhibition of VCAM-1 expression in endothelial cells by CORM-3: the role of the ubiquitin-proteasome system, p38, and mitochondrial respiration.
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.
Fifth Medical Department, University Hospital Mannheim, University of Heidelberg, 68167 Mannheim, Germany.
Claudia
C
Bergstraesser
Simone
S
Hoeger
Hui
H
Song
Linda
L
Ermantraut
Maxi
M
Hottenrot
Tobias
T
Czymai
Marc
M
Schmidt
Matthias
M
Goebeler
Norbert
N
Ponelies
Carsten
C
Stich
Ralf
R
Loesel
Grietje
G
Molema
Marc
M
Seelen
Willem
W
van Son
Benito A
BA
Yard
Neysan
N
Rafat
D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002248:Carbon Monoxide / Q000494:pharmacology; D002478:Cells, Cultured; D018925:Chemokines / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D004789:Enzyme Activation; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D020935:MAP Kinase Signaling System; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D009942:Organometallic Compounds / Q000494:pharmacology*; D010101:Oxygen Consumption; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D059748:Proteolysis; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D057149:Ubiquitinated Proteins / Q000378:metabolism; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism*
D000893:Anti-Inflammatory Agents; D018925:Chemokines; D047908:Intracellular Signaling Peptides and Proteins; D009942:Organometallic Compounds; D014409:Tumor Necrosis Factor-alpha; D057149:Ubiquitinated Proteins; D019010:Vascular Cell Adhesion Molecule-1; C476530:tricarbonylchloro(glycinato)ruthenium(II); D002248:Carbon Monoxide; D046988:Proteasome Endopeptidase Complex