Analysis of new transcription factors induced by stretch force in periodontal ligament cells

牙周膜细胞拉伸力诱导的新转录因子分析

• 批准号: 11671899
• 负责人: MATSUSHITA Kenji
• 金额: $ 2.3万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671899/
• 关键词: periodontal ligament cells; stretch force; CD14; SSRE; NF-kB; proinflammatory cytokine; LPS; 咬合性外傷; 歯周炎; メカニカルストレス; 増殖因子

It is thought that occlusal trauma and dental plaque synergistically contribute to the rapid progression of periodontitis, but we have little information about the underlying mechanism. In this study, I examine the effects of cyclic-stretch force and lipopolysaccharide (LPS) from periodontopathic bacteria on expressions of proinflammatory cytokines in human periodontal ligament cell (HPLC) cultures. Interleukin (IL)-6, IL-8, granulocyte colony stimulating factor (G-CSF), and macrophage colony stimulating factor (M-CSF), and vascular endothelial growth factor (VEGF) were produced by cyclic-stretch force in HPLC cultures. When Prevotella intermedia LPS added to the HPLC cultures simultaneously with or prior to the stimulation with cyclic-stretch force. IL-1α was strongly produced in the HPLC cultures. The productions of IL-6, IL-8, and VEGF were enhanced by the costimulation with cyclic-stretch force and P.intermedia LPS.On the other hand, M-CSF production in HPLC cultures decreased by t … More he costimulation. To clarify molecular mechanism of the enhancement of the induction, we examined expression of CD14, which is a LPS receptor, on stretched HPLC.Although CD14 mRNA was not expressed in non-stretched HPLC, the expression was strongly induced by cyclic-stretch force for 4-8 h in HPLC.Productions of membrane CD14 and soluble CD14 were enhanced by the cyclic-stretch force for 24 h on the surface of and in the supernatants of HPLC respectively. IL-6 production was observed even in the supernatants of stretched and non-stretched HPLC, but the production was enhanced upon serial stimulation with stretch force and P.intermedia LPS on HPLC.The protein kinase C (PKC) inhibitor H7 blocked the expression of CD14 mRNA in stretched HPLC, H7 and the anti-CD14 monoclonal antibody MY4 also suppressed the enhancement of IL-6 production in the HPLC cultures upon serial stimutation with stretch force and P.intermedia LPS.The consensus sequence of shear stress-responsive element (SSRE) was found in the 5' flanking region of the CD14 gene. In addition. a DNA probe which contained the consensus sequence of SSRE bound to nuclear extracts from stretched HPLC but not from non-stretched HPLC.These results suggest that cyclic-stretch force induces CD14 expression in HPLC through PKC activation and that SSRE is associated with the expression. In addition, the expression of CD14 may increase the sensitivity of HPLC to LPS. Less

人们认为咬合创伤和牙菌斑协同促进牙周炎的快速进展，但我们对其潜在机制知之甚少。在这项研究中，我研究了循环拉伸力和牙周病细菌的脂多糖 (LPS) 对人牙周膜细胞 (HPLC) 培养物中促炎细胞因子表达的影响。白细胞介素 (IL)-6、IL-8、粒细胞集落刺激因子 (G-CSF)、巨噬细胞集落刺激因子 (M-CSF) 和血管内皮生长因子 (VEGF) 在 HPLC 培养物中通过循环拉伸力产生。当在循环拉伸力刺激的同时或之前将中间普雷沃氏菌 LPS 添加到 HPLC 培养物中时。 IL-1α 在 HPLC 培养物中大量产生。循环拉伸力和 P.intermedia LPS 的共刺激增强了 IL-6、IL-8 和 VEGF 的产生。另一方面，共刺激 HPLC 培养物中的 M-CSF 产生减少。为了阐明诱导增强的分子机制，我们在拉伸 HPLC 上检测了 LPS 受体 CD14 的表达。虽然 CD14 mRNA 在非拉伸 HPLC 中不表达，但在 HPLC 中循环拉伸力 4-8 小时强烈诱导表达。循环拉伸增强了膜 CD14 和可溶性 CD14 的产量。 分别在 HPLC 表面和上清液中施加 24 小时。即使在拉伸和非拉伸 HPLC 的上清液中也观察到 IL-6 的产生，但在 HPLC 上用拉伸力和中间 P.intermedia LPS 连续刺激后，产生增强。蛋白激酶 C (PKC) 抑制剂 H7 阻断拉伸 HPLC 中 CD14 mRNA 的表达，H7 和抗 CD14 单克隆抗体 MY4 也抑制 IL-6 表达的增强。 在用拉伸力和 P.intermedia LPS 连续刺激后，HPLC 培养物中 IL-6 的产生。在 CD14 基因的 5' 侧翼区域中发现了剪切应力响应元件 (SSRE) 的共有序列。此外。包含与来自拉伸 HPLC 的核提取物结合的 SSRE 共有序列的 DNA 探针，但不与来自非拉伸 HPLC 的核提取物结合。这些结果表明，循环拉伸力通过 PKC 激活诱导 HPLC 中的 CD14 表达，并且 SSRE 与该表达相关。此外，CD14的表达可能会增加HPLC对LPS的敏感性。较少的

项目成果

富田浩一,松下健二,長岡成孝,鳥居光男: "メカニカルストレス・Prevotella intermedia LPS共刺激がヒト歯根膜細胞に与える影響"日本歯科保存学会誌. 42・3. 608-618 (1999)

Koichi Tomita、Kenji Matsushita、Shigetaka Nagaoka、Mitsuo Torii：“机械应力和中间普氏菌 LPS 共刺激对人类牙周膜细胞的影响”日本保守牙科学会杂志 42・3（1999 年）。

Kouichi Tomita, Kenji Matsushita: "Effects of Costimulation with Mechanical Stress and Prevotella intermedia LPS on Human Periodontal Ligament cell. -Induction of Growth Factors and Cytokines-"Japan.J.Conserv.Dent.. 42 (3). 608-618 (1999)

Kouichi Tomita、Kenji Matsushita：“机械应力和中间普氏菌 LPS 共刺激对人牙周膜细胞的影响。-生长因子和细胞因子的诱导-”Japan.J.Conserv.Dent.. 42 (3)。

富田浩一、松下健二 長岡成孝、鳥居光男: "メカニカルストレス・Prevotella intermedia LPS 共刺激がヒト歯根膜細胞に与える影響"日本歯科保存学会誌. 42・3. 608-618 (1999)

Koichi Tomita、Kenji Matsushita、Shigetaka Nagaoka、Mitsuo Torii：“机械应力和中间普氏菌 LPS 共刺激对人类牙周膜细胞的影响”日本保守牙科学会杂志 42・3（1999 年）。