Extracellular human thioredoxin-1 inhibits lipopolysaccharide-induced interleukin-1beta expression in human monocyte-derived macrophages.
Oxidative stress plays an important role in atherosclerotic vascular disease, and several recent studies were focused on thioredoxin-1 (Trx-1) and its potential protective role against oxidative stress. Since human monocyte-derived macrophages (HMDM) are important cells in several inflammatory diseases including atherosclerosis, we conducted this study to evaluate the impact of extracellular recombinant human Trx-1 (rhTrx-1) on gene expression in lipopolysaccharide-activated HMDM. Our results showed that rhTrx-1 was capable of reducing interleukin (IL)-1beta mRNA and protein synthesis in a dose-dependent manner. This effect was partly mediated through a reduction of NF-kappaB activation as analyzed by transient transfection and gel shift assays. In addition, we showed that the attenuation of NF-kappaB activity was the result of the reduction of both p50 and p65 subunit mRNA and protein synthesis on one hand and of the induction of I-kappaBalpha mRNA and protein expression on the other hand. Moreover, inhibition of endogenous Trx-1 mRNA was also observed, suggesting a contribution to the diminution of NF-kappaB activity since endogenous Trx-1, in contrast to the exogenous Trx-1, activates the NF-kappaB system. Finally, H2O2-oxidized rhTrx-1 reduced IL-1beta mRNA synthesis in lipopolysaccharide-activated HMDM. This result highly suggested that the rhTrx-1 used in this study could be oxidized in the culture medium and, in turn, reduced IL-1beta mRNA and protein synthesis. Taken together, these data indicated a potential new mechanism through which extracellular rhTrx-1 exerts an anti-inflammatory function in HMDM.
Billiet
Ludivine
L
U-545 INSERM, Institut Pasteur de Lille and Université Lille 2, 59019 Lille, France.
Furman
Christophe
C
Larigauderie
Guilhem
G
Copin
Corinne
C
Brand
Korbinian
K
Fruchart
Jean-Charles
JC
Rouis
Mustapha
M
D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D050197:Atherosclerosis; D015153:Blotting, Western; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D020672:I-kappa B Proteins / Q000378:metabolism; D007249:Inflammation; D007375:Interleukin-1 / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism*; D008954:Models, Biological; D009000:Monocytes / Q000378:metabolism*; D000072000:NF-KappaB Inhibitor alpha; D052002:NF-kappa B p50 Subunit / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000737:chemistry; D018384:Oxidative Stress; D011401:Promoter Regions, Genetic; D012313:RNA / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013879:Thioredoxins / Q000378:metabolism*; D051996:Transcription Factor RelA / Q000378:metabolism; D014162:Transfection
D000893:Anti-Inflammatory Agents; D020672:I-kappa B Proteins; D007375:Interleukin-1; D008070:Lipopolysaccharides; D052002:NF-kappa B p50 Subunit; C000606204:NFKBIA protein, human; D009841:Oligonucleotides; D012333:RNA, Messenger; D011994:Recombinant Proteins; D051996:Transcription Factor RelA; D000072000:NF-KappaB Inhibitor alpha; D013879:Thioredoxins; D012313:RNA; D006861:Hydrogen Peroxide