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Prostaglandin E₂ Regulates Interleukin-1β-induced Matrix Metalloproteinase-3 Production in Human Gingival Fibroblasts

¹ Periodontology, Department of Hard Tissue Engineering, Graduate School, and ² Centre of Excellence Program for Frontier Research on Molecular Destruction of Tooth and Bone, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8549, Japan;

View larger version (18K): [in this window] [in a new window]   Figure 1. Effect of indomethacin on MMP-3 (A,B) and PGE2 (C) production in IL-1β-stimulated HGF derived from healthy gingiva. These cells were stimulated with vehicle or 2 ng/mL IL-1β in the presence or absence of 1 µM of indomethacin for 72 hrs. After incubation, MMP-3 (A) and PGE2 levels (C) and caseinolytic activities (B) in the culture media were evaluated by enzyme-linked immunosorbent assay and casein zymography, as described in MATERIALS & METHODS. Values are mean + SD (n = 4). Data are representative of 3 separate experiments. *Significantly different from control (p < 0.0001). **Significantly different from IL-1β alone (p < 0.001). #Significantly different from control (p < 0.0001). ##Significantly different from IL-1β alone (p < 0.001).

View larger version (52K): [in this window] [in a new window]   Figure 2. Effects of PGE2, various EP agonists, or cAMP-elevating agents on MMP-3 levels and activities in IL-1β-stimulated HGF derived from healthy gingiva. Cells were stimulated with vehicle or 2 ng/mL IL-1β in the presence of 1 µM indomethacin, with or without various doses of PGE2, 17 phenyl--trinor PGE2, butaprost, and ONO-AE-1329 for caseinolytic activity assay (A,B,C,D), or with or without various doses of PGE2, 1 µM of 17 phenyl--trinor PGE2, 1 µM of butaprost, 1 µM of ONO-AE1-329, 100 µM of dibutyryl cAMP (dbcAMP), and 10 µM of forskolin (FS) for MMP-3 protein level assay (E,F,G,H), for 72 hrs. After incubation, caseinolytic activities and MMP-3 protein levels in the culture media were assayed by casein zymography and enzyme-linked immunosorbent assay, respectively, as described in MATERIALS & METHODS. Values are mean ± SD (n = 4). Data are representative of 3 separate experiments. *Significantly different from indomethacin+1L-1β (p < 0.05). **Significantly different from indomethacin+1L-1β (p < 0.0001). ***Significantly different from indomethacin+1L-1β (p < 0.001). #Significantly different from indomethacin+1L-1β (p < 0.05). $Significantly different from indomethacin+1L-1β (p < 0.05). $$Significantly different from indomethacin+1L-1β (p < 0.005). @Significantly different from indomethacin+1L-1β (p < 0.0001).

View larger version (25K): [in this window] [in a new window]   Figure 3. Effect of indomethacin on MMP-3 (A,B) and PGE2 (C) production in IL-1β-stimulated HGF derived from periodontally affected tissue. Cells were stimulated with vehicle or 2 ng/mL IL-1β in the presence or absence of 1 µM of indomethacin for 72 hrs. After incubation, MMP-3 (A) and PGE2 (C) levels and caseinolytic activities (B) in the culture media were evaluated by enzyme-linked immunosorbent assay and casein zymography, as described in MATERIALS & METHODS. Values are mean ± SD (n = 4). Data are representative of 3 separate experiments. *Significantly different from control (p < 0.0001). **Significantly different from IL-1β alone (p < 0.001). #Significantly different from control (p < 0.0001). ##Significantly different from IL-1β alone (p < 0.001).

View larger version (23K): [in this window] [in a new window]   Figure 4. Effects of PGE2, 17 phenyl--trinor PGE2, butaprost, and ONO-AE1-329 on production of MMP-3 in IL-1β-stimulated HGF derived from periodontally affected gingiva. Cells were stimulated with vehicle or 2 ng/mL IL-1β in the presence of 1 µM indomethacin, with or without various doses of PGE2, 17 phenyl--trinor PGE2, butaprost, and ONO-AE1-329 for caseinolytic activity assay (A,B,C,D), or with or without 1 µM of PGE2, 17 phenyl--trinor PGE2, butaprost, and ONO-AE1-329 for MMP-3 protein level assay (E), for 72 hrs. After incubation, caseinolytic activities and MMP-3 protein levels in the culture media were assayed by casein zymography and enzyme-linked immunosorbent assay, respectively, as described in MATERIALS & METHODS. Values are mean ± SD (n = 4). Data are representative of 3 separate experiments. *Significantly different from indomethacin+IL-1β (p < 0.0001).

Journal of Dental Research, Vol. 83, No. 3, 260-265 (2004)
DOI: 10.1177/154405910408300315


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