Effects of JNK1/2 on the inflammation cytokine TNF-α-enhanced production of MMP-3 in human dental pulp fibroblast-like cells.
OBJECTIVE
To investigate the effects of the c-Jun N-terminal kinase (JNK1/2) on the inflammation cytokine tumour necrosis factor-alpha (TNF-α)-enhanced production of matrix metalloproteinase-3 (MMP-3) in human dental pulp fibroblast-like cells (HPFs).


METHODS
HPFs were grown from pulp explants from healthy donors. Primary cultures were established by culturing the cells for 20 to 30 days. The experiments with HPFs were performed between passages 3 and 10. The HPFs were incubated in serum-free medium containing TNF-α for 24 h. The medium in each well was prepared in SDS sample buffer and was analysed for MMP-3 by Western blotting.


RESULTS
JNK inhibitor SP601245 markedly inhibited the production of MMP-3 in TNF-α-stimulated human dental pulp fibroblasts. MMP-3 production was enhanced by TNF-α in HPFs; silencing JNK1 and JNK2 expression inhibited this activation. cAMP response element-binding protein (CREB) was activated by TNF-α in HPFs; silencing JNK1 and JNK2 expression inhibited this activation.


CONCLUSIONS
The activation of CREB via JNK pathways in the presence of TNF-α occurred with enhancement of MMP-3 production in dental pulp fibroblasts.
Department of Biochemistry, Osaka Dental University, Hirakata, Japan.
S
S
Goda
Department of Endodontics, Osaka Dental University, Hirakata, Japan.
Y
Y
Kato
Department of Biochemistry, Osaka Dental University, Hirakata, Japan.
E
E
Domae
Department of Endodontics, Osaka Dental University, Hirakata, Japan.
H
H
Hayashi
Department of Pulp Biology and Endodontics, Kanagawa Dental University, Yokosuka, Japan.
N
N
Tani-Ishii
Department of Cell Biology, Windber Research Institute, Windber, USA.
J
J
Iida
Department of Biochemistry, Osaka Dental University, Hirakata, Japan.
T
T
Ikeo
D015153:Blotting, Western; D002478:Cells, Cultured; D003782:Dental Pulp / Q000166:cytology*; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D014409:Tumor Necrosis Factor-alpha; D048031:JNK Mitogen-Activated Protein Kinases; C511768:MMP3 protein, human; D019278:Matrix Metalloproteinase 3
C-Jun N-terminal kinase; human dental pulp fibroblast-like cells; matrix metalloproteinase-3