Amelioration of acute inflammation by systemic administration of a cell-permeable peptide inhibitor of NF-kappaB activation.
OBJECTIVE
We used an experimental model of inflammation in mice, carrageenan-induced paw edema, to study the antiinflammatory effects of the NEMO-binding domain (NBD) peptide, which blocks activation of the inducible transcription factor NF-kappaB.


METHODS
Paw edema was induced by subplantar injection of 1% lambda-carrageenan into the mouse left hind paw. Test agents were given intraperitoneally immediately after carrageenan injection. The increase in footpad thickness was considered to be edema. In some experiments, the mice were killed and the paws were removed for histologic and molecular biology analysis. NF-kappaB DNA binding activity was evaluated in nuclear extracts by electrophoretic mobility shift assays. The expression levels of NF-kappaB-regulated cyclooxygenase 2 (COX-2) protein and tumor necrosis factor alpha (TNFalpha) messenger RNA (mRNA) were evaluated by immunoblot analysis and polymerase chain reaction amplification of reverse-transcribed mRNA, respectively.


RESULTS
We found that systemically administered NBD peptide significantly inhibited edema formation and cellular infiltration in inflamed mouse paws. This antiinflammatory activity was most likely due to inhibition of expression of proinflammatory mediators, such as TNFalpha and COX-2, in inflamed tissues.


CONCLUSIONS
These studies further establish NF-kappaB as a target for antiinflammatory therapy and provide support for the use of the NBD peptide as a possible therapeutic agent for inflammatory diseases.
di Meglio
Paola
P
Yale University School of Medicine, New Haven, Connecticut 06520, USA.
Ianaro
Angela
A
Ghosh
Sankar
S
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000276:immunology / Q000627:therapeutic use*; D002351:Carrageenan / Q000009:adverse effects; D051546:Cyclooxygenase 2; D004487:Edema / Q000139:chemically induced / Q000188:drug therapy* / Q000276:immunology; D005260:Female; D006614:Hindlimb / Q000276:immunology; D007249:Inflammation / Q000139:chemically induced / Q000188:drug therapy* / Q000276:immunology; D051379:Mice; D023421:Models, Animal; D016328:NF-kappa B / Q000276:immunology*; D010455:Peptides / Q000276:immunology / Q000627:therapeutic use*; D011134:Polysaccharides / Q000009:adverse effects; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D000893:Anti-Inflammatory Agents; C467063:NBD peptide, mouse; D016328:NF-kappa B; D010455:Peptides; D011134:Polysaccharides; D014409:Tumor Necrosis Factor-alpha; D002351:Carrageenan; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases