ヒト歯髄由来線維芽細胞におけるIL-1β刺激によるMMP-3産生の影響
目的 : 歯髄は, 象牙質への栄養供給・恒常性の維持などにかかわり, 修復象牙質形成に対し多大な役割を有している. 一方で, 齲蝕の進行に伴う歯髄の炎症によって歯髄組織にはInterleukin-1β (IL-1β) などの炎症性サイトカインが産生され, Matrix metalloproteinases (MMPs) などの酵素が産生され組織破壊へと進行していく. 歯髄炎の発症機序を解明することは, 歯髄保存のために重要である. そこでわれわれは, ヒト歯髄由来線維芽細胞における炎症性サイトカインIL-1β刺激によるMMP-3の産生と, 細胞内シグナル伝達物質であるExtracellular...
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| Published in | 日本歯科保存学雑誌 Vol. 57; no. 1; pp. 1 - 8 |
|---|---|
| Main Authors | , , , , , |
| Format | Journal Article |
| Language | Japanese |
| Published |
特定非営利活動法人 日本歯科保存学会
2014
日本歯科保存学会 |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0387-2343 2188-0808 |
| DOI | 10.11471/shikahozon.57.1 |
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| Abstract | 目的 : 歯髄は, 象牙質への栄養供給・恒常性の維持などにかかわり, 修復象牙質形成に対し多大な役割を有している. 一方で, 齲蝕の進行に伴う歯髄の炎症によって歯髄組織にはInterleukin-1β (IL-1β) などの炎症性サイトカインが産生され, Matrix metalloproteinases (MMPs) などの酵素が産生され組織破壊へと進行していく. 歯髄炎の発症機序を解明することは, 歯髄保存のために重要である. そこでわれわれは, ヒト歯髄由来線維芽細胞における炎症性サイトカインIL-1β刺激によるMMP-3の産生と, 細胞内シグナル伝達物質であるExtracellular signal-regulated kinase 1/2 (ERK1/2) について検討した. 材料と方法 : 大阪歯科大学医の倫理委員会における承認 (大歯医倫070716号) の下, 矯正治療における便宜抜去歯からヒト歯髄由来線維芽細胞を採取し, 初代培養後3∼10世代を本研究に供した. ヒト歯髄由来線維芽細胞はα-MEM (serum-free) にて培養後, IL-1β (0, 1, 2, 5, 10ng/ml) およびMEK阻害剤 (U0126) を添加した. 刺激後のMMP-3の産生およびERK1/2のリン酸化を, それぞれの抗体を用いてウエスタンブロット法にて検討した. また, 上清を1mg/mlのゼラチンを加えたSDS-PAGEにて電気泳動を行い, ゼラチンザイモグラフィー法にて検討した. 成績 : ヒト歯髄由来線維芽細胞において, IL-1β刺激によりMMP-3の産生は増強した. ERK1/2のリン酸化は, IL-1β濃度依存性に増強した. さらに, MEK1/2阻害剤U0126添加することで, IL-1β刺激により増強したERK1/2のリン酸化およびMMP-3の産生が阻害された. 考察 : 本実験の結果から, ヒト歯髄由来線維芽細胞におけるIL-1β刺激によるMMP-3産生はMEK1/2・ERK1/2経路が関与していることが示唆された. |
|---|---|
| AbstractList | 「抄録」 目的:歯髄は, 象牙質への栄養供給・恒常性の維持などにかかわり, 修復象牙質形成に対し多大な役割を有している. 一方で, 齲蝕の進行に伴う歯髄の炎症によって歯髄組織にはInterleukin-1β(IL-1β)などの炎症性サイトカインが産生され, Matrix metalloproteinases(MMPs)などの酵素が産生され組織破壊へと進行していく. 歯髄炎の発症機序を解明することは, 歯髄保存のために重要である. そこでわれわれは, ヒト歯髄由来線維芽細胞における炎症性サイトカインIL-1β刺激によるMMP-3の産生と, 細胞内シグナル伝達物質であるExtracellular signal-regulated kinase 1/2(ERK1/2)について検討した. 材料と方法:大阪歯科大学医の倫理委員会における承認(大歯医倫070716号)の下, 矯正治療における便宜抜去歯からヒト歯髄由来線維芽細胞を採取し, 初代培養後3～10世代を本研究に供した. ヒト歯髄由来線維芽細胞はα-MEM(serum-free)にて培養後, IL-1β(0, 1, 2, 5, 10ng/ml)およびMEK阻害剤(U0126)を添加した. 刺激後のMMP-3の産生およびERK1/2のリン酸化を, それぞれの抗体を用いてウエスタンブロット法にて検討した. また, 上清を1mg/mlのゼラチンを加えたSDS-PAGEにて電気泳動を行い, ゼラチンザイモグラフィー法にて検討した. 成績:ヒト歯髄由来線維芽細胞において, IL-1β刺激によりMMP-3の産生は増強した. ERK1/2のリン酸化は, IL-1β濃度依存性に増強した. さらに, MEK1/2阻害剤U0126添加することで, IL-1β刺激により増強したERK1/2のリン酸化およびMMP-3の産生が阻害された. 考察:本実験の結果から, ヒト歯髄由来線維芽細胞におけるIL-1β刺激によるMMP-3産生はMEK1/2・ERK1/2経路が関与していることが示唆された. 目的 : 歯髄は, 象牙質への栄養供給・恒常性の維持などにかかわり, 修復象牙質形成に対し多大な役割を有している. 一方で, 齲蝕の進行に伴う歯髄の炎症によって歯髄組織にはInterleukin-1β (IL-1β) などの炎症性サイトカインが産生され, Matrix metalloproteinases (MMPs) などの酵素が産生され組織破壊へと進行していく. 歯髄炎の発症機序を解明することは, 歯髄保存のために重要である. そこでわれわれは, ヒト歯髄由来線維芽細胞における炎症性サイトカインIL-1β刺激によるMMP-3の産生と, 細胞内シグナル伝達物質であるExtracellular signal-regulated kinase 1/2 (ERK1/2) について検討した. 材料と方法 : 大阪歯科大学医の倫理委員会における承認 (大歯医倫070716号) の下, 矯正治療における便宜抜去歯からヒト歯髄由来線維芽細胞を採取し, 初代培養後3∼10世代を本研究に供した. ヒト歯髄由来線維芽細胞はα-MEM (serum-free) にて培養後, IL-1β (0, 1, 2, 5, 10ng/ml) およびMEK阻害剤 (U0126) を添加した. 刺激後のMMP-3の産生およびERK1/2のリン酸化を, それぞれの抗体を用いてウエスタンブロット法にて検討した. また, 上清を1mg/mlのゼラチンを加えたSDS-PAGEにて電気泳動を行い, ゼラチンザイモグラフィー法にて検討した. 成績 : ヒト歯髄由来線維芽細胞において, IL-1β刺激によりMMP-3の産生は増強した. ERK1/2のリン酸化は, IL-1β濃度依存性に増強した. さらに, MEK1/2阻害剤U0126添加することで, IL-1β刺激により増強したERK1/2のリン酸化およびMMP-3の産生が阻害された. 考察 : 本実験の結果から, ヒト歯髄由来線維芽細胞におけるIL-1β刺激によるMMP-3産生はMEK1/2・ERK1/2経路が関与していることが示唆された. |
| Author | 堂前, 英資 竹内, 摂 吉川, 一志 池尾, 隆 合田, 征司 山本, 一世 |
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J Dent Res 2009; 88: 339-344. 17) Zhai S, Wang Y, Jiang W, Jia Q, Li J, Wang W, Wang H, Ding Y, Wang P, Liu J, Ni L. Nemotic human dental pulp fibroblasts promote human dental pulp stem cells migration. Exp Cell Res 2013; 319: 1544-1552. 23) Sorsa T, Tjaderhane L, Salo T. Matrix metalloproteinases (MMPs) in oral diseases. Oral Dis 2004; 10: 311-318. |
| References_xml | – reference: 29) Buonomo R, Giacco F, Vasaturo A, Caserta S, Guido S, Pagliara V, Garbi C, Mansueto G, Cassese A, Perruolo G, Oriente F, Miele C, Beguinot F, Formisano P. PED/PEA-15 controls fibroblast motility and wound closure by ERK1/2-dependent mechanisms. J Cell Physiol 2012; 227: 2106-2116. – reference: 33) Wisithphrom K, Murray PE, Windsor LJ. Interleukin-1 alpha alters the expression of matrix metalloproteinases and collagen degradation by pulp fibroblasts. J Endod 2006; 32: 186-192. – reference: 5) Kitamura C, Nishihara T, Ueno Y, Nagayoshi M, Kasugai S, Terashita M. Thermotolerance of pulp cells and phagocytosis of apoptotic pulp cells by surviving pulp cells following heat stress. J Cell Biochem 2005; 94: 826-834. – reference: 22) Ozeki N, Yamaguchi H, Kawai R, Hiyama T, Nakata K, Mogi M, Nakamura H. Cytokines induce MMP-3-regulated proliferation of embryonic stem cell-derived odontoblast-like cells. 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| Snippet | 目的 : 歯髄は, 象牙質への栄養供給・恒常性の維持などにかかわり, 修復象牙質形成に対し多大な役割を有している. 一方で, 齲蝕の進行に伴う歯髄の炎症によって歯髄組織にはInterleukin-1β (IL-1β) などの炎症性サイトカインが産生され, Matrix metalloproteinases... 「抄録」 目的:歯髄は, 象牙質への栄養供給・恒常性の維持などにかかわり, 修復象牙質形成に対し多大な役割を有している. 一方で, 齲蝕の進行に伴う歯髄の炎症によって歯髄組織にはInterleukin-1β(IL-1β)などの炎症性サイトカインが産生され, Matrix... |
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| SubjectTerms | ERK1/2 MAP kinase Interleukin-1β MMPs ヒト歯髄由来線維芽細胞 |
| Title | ヒト歯髄由来線維芽細胞におけるIL-1β刺激によるMMP-3産生の影響 |
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