Antioxidants inhibit tumor necrosis factor-alpha mediated stimulation of interleukin-8, monocyte chemoattractant protein-1, and collagenase expression in cultured human synovial cells.
OBJECTIVE
To study whether the induction of mRNA for interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and collagenase by tumor necrosis factor-alpha (TNF-alpha) is suppressed by antioxidants in human synovial cells. TNF-alpha has been shown to exert some of its effects by stimulating production of reactive oxygen intermediates in some cell lines other than synovial cells.


METHODS
Amounts of mRNA for IL-8, MCP-1, and collagenase were determined by Northern blot analysis. Electrophoretic mobility shift assays were performed for the detection of a transcription factor, nuclear factor-kappa B(NF-kappa B). The concentration of IL-8 in the medium was determined by ELISA.


RESULTS
TNF-alpha increased the expression of IL-8, MCP-1, and collagenase mRNA in human synovial cells. NF-KB known to induce IL-8 gene transcription was also increased in nuclear extracts from the synovial cells treated with TNF-alpha. Prior addition of antioxidant, N-acetyl-L-cysteine (NAC) or 2-oxothiazolidine-4-carboxylate (OTC), suppressed TNF-alpha stimulated expressions of IL-8, MCP-1, and collagenase mRNA in a dose dependent manner. Treatment with NAC also suppressed TNF-alpha induced increase in NF-kappa B. The changes of IL-8 in the medium reflected the mRNA levels. Hydrogen peroxide (H2O2) induced the expression of mRNA for the cytokines but not collagenase mRNA, and NAC suppressed the effect of H2O2.


CONCLUSIONS
Our data suggest that TNF-alpha induces expression of proinflammatory cytokines such as IL-8 and MCP-1 through generation of reactive oxygen intermediates and subsequent activation of NF-kappa B in human synovial cells, and the antioxidants may inhibit, at least in part, the activation of NF-kappa B by TNF-alpha. These results indicate that antioxidants such as NAC may be useful in treating rheumatoid arthritis.
Sato
M
M
Department of Endocrinology and Metabolism Research, Nagoya University, Furo-cho Chikusa-ku, Japan.
Miyazaki
T
T
Nagaya
T
T
Murata
Y
Y
Ida
N
N
Maeda
K
K
Seo
H
H
D000111:Acetylcysteine / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology*; D001172:Arthritis, Rheumatoid / Q000188:drug therapy; D002478:Cells, Cultured / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology; D018932:Chemokine CCL2 / Q000235:genetics*; D017364:Collagenases / Q000235:genetics*; D016166:Free Radical Scavengers / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000276:immunology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D016209:Interleukin-8 / Q000235:genetics*; D016328:NF-kappa B / Q000235:genetics; D011761:Pyrrolidonecarboxylic Acid; D012333:RNA, Messenger / Q000378:metabolism; D013583:Synovial Membrane / Q000166:cytology; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines; D014158:Transcription, Genetic / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D000975:Antioxidants; D018932:Chemokine CCL2; D016166:Free Radical Scavengers; D016209:Interleukin-8; D016328:NF-kappa B; D012333:RNA, Messenger; D013844:Thiazoles; D053778:Thiazolidines; D014409:Tumor Necrosis Factor-alpha; D006861:Hydrogen Peroxide; D017364:Collagenases; D011761:Pyrrolidonecarboxylic Acid; D000111:Acetylcysteine; C029536:2-oxothiazolidine-4-carboxylic acid