Association of interleukin-1-induced, NF kappa B DNA-binding activity with collagenase gene expression in human gingival fibroblasts.
During earlier examination of interleukin-1 (IL-1)-induced matrix metalloproteinase gene expression in human gingival fibroblasts a highly induced immediate early gene, I kappa B-alpha, a NF kappa B DNA-binding inhibitor, was identified. The aim now was to investigate whether recombinant (r)IL-1 beta induces the stimulation of NF kappa B and its inhibitor proteins in human gingival fibroblasts and to understand if inhibition of its activity affects collagenase gene expression. Primary gingival fibroblasts (human) were treated with rIL-1 beta to determine the effect on NF kappa B-like DNA-binding activity. IL-1 induced the production of steady-state mRNA levels of I kappa B-alpha in the cultured fibroblasts. Nuclear run-on transcription studies demonstrated that rIL-1 induction of I kappa B-alpha may be transcriptionally regulated. Using electrophoretic mobility gel-shift assays it was shown that rIL-1 activates NF kappa B-like, DNA-binding activity in these fibroblasts. NF kappa B-like DNA-binding activity was rapidly induced and turned over in gingival fibroblasts with peak activity at 30 min after rIL-1 treatment. Further, treatment with chymotrypsin protease inhibitor and antioxidant inhibitor prevented IL-1-induced, NF kappa B-like, DNA-binding activity and collagenase mRNA production. When coupled with the existence of NF kappa B consensus DNA-binding sites on the collagenase gene promoter, these findings suggest that the stimulation of NF kappa B in gingival fibroblasts by rIL-1 could play an important part in the regulation of their collagenase gene expression. The ability of IL-1 to stimulate this expression may define a pivotal role for this cytokine in the pathogenesis of periodontitis.
Tewari
M
M
Department of Orthodontics, School of Dentistry, Temple University, Philadelphia, PA 19140, USA.
Tuncay
O C
OC
Milchman
A
A
Reddy
P J
PJ
Reddy
C D
CD
Cressman
D E
DE
Taub
R
R
Newton
R C
RC
Tewari
D S
DS
D002478:Cells, Cultured; D002874:Chromosome Mapping; D002918:Chymotrypsin / Q000037:antagonists & inhibitors; D017364:Collagenases / Q000235:genetics*; D016384:Consensus Sequence / Q000235:genetics; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D017781:Genes, Immediate-Early / Q000235:genetics; D005881:Gingiva / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology; D010518:Periodontitis / Q000209:etiology / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins; D014158:Transcription, Genetic / Q000235:genetics
D007375:Interleukin-1; D016328:NF-kappa B; D016877:Oxidants; D012333:RNA, Messenger; D011994:Recombinant Proteins; D004247:DNA; D002918:Chymotrypsin; D017364:Collagenases