Effect of transmembrane proteoglycans in human periodontal ligament fibroblasts

跨膜蛋白多糖对人牙周膜成纤维细胞的影响

• 批准号: 13672154
• 负责人: SHIMAZU Atsushi
• 金额: $ 2.3万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672154/
• 关键词: periodontal ligament; basic fibroblast growth factor; matrix metalloproteinase (MMP)-3; 線維芽細胞成長因子; MMP-3; プロテオグリカン

Basic fibroblast growth factor (bFGF, FGF-2) is one of the potent mitogens for periodontal ligament (PDL) cells. However, the role of bFGF on the matrix metalloproteinase (MMP) -3 expression in PDL cells is unknown. In this study, the effect of bFGF on MMP-3 expression in PDL cells and the mechanism of this process were examined.Human PDL cells were exposed to bFGF at various concentrations (0.01 - 10 ng/ml) in monolayer cultures. bFGF increased [3H]-thymidine incorporation and suppressed proteoglycan synthesis concentration-dependently. However, similar concentration ranges of bFGF increased the release of the cell-associated proteoglycans into the medium. Furthermore, bFGF increased MMP-3 mRNA levels concentration-dependently as examined by reverse transcription-polymerase chain reaction. Induction of MMP-3 after the stimulation with bFGF was observed as early as 12 h with maximal at 24 h. Thereafter the MMP-3 mRNA level gradually decreased until 72 h. Cycloheximide blocked the induction of MMP-3 by bFGF, indicating the requirement of de novo protein synthesis for this stimulation. Furthermore, MMP-3 expression induced by bFGF was abrogated by U0126, a specific inhibitor of MEK1/2 and ERK1/2 in mitogen-activated protein (MAP) kinase pathway, not by PD98059, a specific inhibitor of MEK1. In addition, bFGF up-regulated the phosphorylated ERK1/2 in 5 min with the maximal at 20 min as examined by Western blotting, and U0126 inhibited the ERK1/2 phosphorylation induced by bFGF.These findings suggest that bFGF induces MMP-3 expression in PDL cells through the activation of the MEK2 in MAP kinase pathway. bFGF stimulation on MMP-3 synthesis may be involved in the control of the cell-associated proteoglycans in PDL cells during periodontal regeneration and degradation.

碱性成纤维细胞生长因子（bFGF, FGF-2）是牙周韧带（PDL）细胞的有效丝裂原之一。然而，bFGF在PDL细胞中基质金属蛋白酶(MMP) -3表达中的作用尚不清楚。本研究探讨bFGF对PDL细胞MMP-3表达的影响及其作用机制。将人PDL细胞暴露于不同浓度（0.01 - 10 ng/ml）的bFGF单层培养物中。bFGF增加[3H]-胸腺嘧啶结合，抑制蛋白多糖合成，呈浓度依赖性。然而，相似浓度范围的bFGF增加了细胞相关蛋白聚糖向培养基中的释放。此外，通过逆转录聚合酶链反应检测，bFGF增加MMP-3 mRNA水平呈浓度依赖性。bFGF刺激后MMP-3的诱导最早在12 h， 24 h达到峰值，此后MMP-3 mRNA水平逐渐下降，直到72 h。环己亚胺阻断了bFGF对MMP-3的诱导，表明这种刺激需要从头合成蛋白质。此外，由bFGF诱导的MMP-3表达被丝裂原活化蛋白（MAP）激酶通路中MEK1/2和ERK1/2特异性抑制剂U0126所抑制，而不是MEK1特异性抑制剂PD98059所抑制。Western blotting检测bFGF在5 min内上调磷酸化的ERK1/2, 20 min上调幅度最大，U0126抑制bFGF诱导的ERK1/2磷酸化。这些结果表明，bFGF通过激活MAP激酶通路中的MEK2诱导PDL细胞中MMP-3的表达。bFGF对MMP-3合成的刺激可能参与了牙周再生和降解过程中PDL细胞相关蛋白聚糖的控制。

项目成果

Yamasaki, Y. et al.: "Synthesis of functionally graded MgCO3 apatite accelerating osteoblast adhesion"Journal of Biomedical Materials Research. (in press).

Yamasaki, Y. 等人：“加速成骨细胞粘附的功能分级 MgCO3 磷灰石的合成”生物医学材料研究杂志。

Shimazu, A. et al.: "Basic fibroblast growth factor induces the expression of matrix metalloproteinase-in human periodontal ligament cells through the MEK2 mitogen-activated protein kinase pathway"Journal of Periodontal Research. 38. 122-129 (2003)

Shimazu, A. 等人：“碱性成纤维细胞生长因子通过 MEK2 丝裂原激活蛋白激酶途径诱导人牙周膜细胞中基质金属蛋白酶的表达”《牙周研究杂志》。

島津 篤 他: "粘膜ワクチン、経皮ワクチン研究の現状"最新医学. 57. 71-78 (2002)

Atsushi Shimazu 等：“粘膜和透皮疫苗研究的现状”现代医学 57. 71-78 (2002)。

Tsutsumi, S. et al.: "Retention of Multi-lineage differentiation potential of mesenchymal cells during proliferation in response to FGF."Biochemical and Biophysical Research Communications. 288. 413-419 (2001)

Tsutsumi, S. 等人：“在响应 FGF 的增殖过程中保留间充质细胞的多谱系分化潜力。”生物化学和生物物理研究通讯。

Tsutsumi, S. et al.: "Retention of Multi-lineage differentiation potential of mesenchymal cells during proliferation in response to FGF"Biochemical and Biophysical Research Communications. 288. 413-419 (2001)

Tsutsumi, S. 等人：“在响应 FGF 的增殖过程中保留间充质细胞的多谱系分化潜力”生物化学和生物物理研究通讯。