Inhibition of TNF-alpha, induced cytokine and adhesion molecule. Expression in glomerular cells in vitro and in vivo by transcription factor decoy for NFkappaB.
Activation of certain cytokines and adhesion molecules has been postulated being involved in the pathogenesis of experimental and human glomerulonephritis. In this study, we examined whether the transcription factor, nuclear factor kappaB (NFkappaB), mediated the expression of these genes involved with the inflammatory response of mesangial cells by using transcription factor decoy oligodeoxynucleotides (ODN) to block NFkappaB binding to the promoter site of its target genes. We hypothesized that the NFkappaB decoy ODN can inhibit the coordinated activation of cytokines and adhesion molecules induced by TNF-alpha. Increased binding activity of NFkappaB induced by TNF-alpha was effectively blocked by the NFkappaB decoy ODN. TNF-alpha stimulated CAT expression, which was significantly inhibited by transfection of NFkappaB, but not by scrambled decoy ODN. Of importance, NFkappaB, but not scrambled decoy ODN, significantly attenuated the increase in RNA and protein levels of IL-1alpha, IL-1beta, IL-6, ICAM-1 and VCAM-1 induced by TNF-alpha assessed by RT-PCR. Moreover, in vivo transfection of NFkappaB decoy ODN inhibited expressions of these cytokines and adhesion molecules induced by TNF-alpha injection. These results suggest a novel therapeutic strategy for the treatment of glomerulonephritis using decoy ODN to block the binding of NFkappaB, inhibiting the coordinated transactivation of the key cytokines and adhesion molecules, and thereby suppressing the inflammatory process.
Tomita
N
N
Department of General Medicine, Osaka University Hospital, Suita, Japan. tomita@hp-gm.med.osaka-u.ac.jp
Morishita
R
R
Tomita
S
S
Kaneda
Y
Y
Higaki
J
J
Ogihara
T
T
Horiuchi
M
M
D000818:Animals; D001665:Binding Sites; D015815:Cell Adhesion Molecules / Q000096:biosynthesis*; D002461:Cell Line, Transformed; D016207:Cytokines / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D005920:Glomerular Mesangium / Q000378:metabolism*; D066298:In Vitro Techniques; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D008081:Liposomes; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D009841:Oligonucleotides / Q000235:genetics; D018108:Respirovirus / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
D015815:Cell Adhesion Molecules; D016207:Cytokines; D008081:Liposomes; D016328:NF-kappa B; D009841:Oligonucleotides; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1