PERIO - Periodontal Practice Today 5 (2008), No. 3 (18.11.2008)
Effect of nuclear factor-kB inhibition on interleukin-1ß-stimulated matrix metalloproteinase-3 production by gingival fibroblasts from a patient with aggressive periodontitis
Tipton, David A. / Pond, David M.
Background: Aggressive periodontitis (AgP) causes rapid periodontal destruction. Matrix metalloproteinase- 3 (MMP-3) degrades the extracellular matrix and is associated with periodontitis. Interleukin-1ß (IL-1ß) upregulates MMP-3 in human gingival fibroblasts, activating genes via transcription factors (i.e. nuclear factor ΚB [NF-ΚB]). Cytoplasmic NF-ΚB is bound to inhibitors of KB (IKB), which are phosphorylated and degraded after cellular stimulation by IL-1ß, allowing nuclear translocation of NF-ΚB subunits (e.g. p65). This involves activation of IΚB kinase (IKK), requiring a regulatory subunit [NF-ΚB essential modifier (NEMO)]. The objective of this study was to investigate NF-ΚB involvement in IL- 1ß upregulation of MMP-3 in AgP gingival fibroblasts by inhibiting NF-ΚB activation with NEMO binding domain (NBD) peptide, blocking NEMO-IKK binding.
Study design: Constitutive and IL-1ß-stimulated MMP-3 was measured by enzyme-linked immunosorbent assay (ELISA). For effects on NF-ΚB activation, cells were preincubated with 50 to 200µM NBD or control peptide (CP) for 6 to 48h, stimulated with IL-1ß, and nuclear p65 levels were measured. For effects on MMP-3, cells were preincubated with 50µM NBD or CP for 24 or 48h, stimulated with IL-1ß, and MMP-3 was measured.
Results: There were time-dependent increases in constitutive MMP-3 production, and IL-1ß significantly increased MMP-3 at each time point. NBD peptide specifically decreased IL-1ß-stimulated nuclear p65 by approximately 55%, and decreased IL-1ß-stimulated MMP-3 production by a maximum of 26%.
Conclusions: IL-1ß stimulation of MMP-3 may be partially regulated by NF-ΚB. Blocking MMP-3 gene expression by inhibitors such as NBD peptide may be an important approach for treatment of AgP.
Keywords: aggressive periodontitis, IL-1ß, MMP-3, NBD peptide, NF-ΚB