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

人们认为咬合创伤和牙菌斑协同作用导致牙周炎的快速进展，但我们对潜在机制的信息很少。在这项研究中，我检查周期性的拉伸力和脂多糖（LPS）牙周病细菌的促炎细胞因子在人牙周膜细胞（HPLC）培养物的表达的影响。在HPLC培养物中通过循环拉伸力产生白细胞介素（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）。当中间普雷沃氏菌LPS加入到HPLC培养物中的同时或之前，用循环拉伸力刺激。在HPLC培养物中强烈产生IL-1α。结果表明，周期性牵张力和LPS共刺激可促进IL-6、IL-8和VEGF的产生，而M-CSF的产生则因周期性牵张力和LPS的共同刺激而减少。 ...更多信息 共刺激。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CD 14的表达。虽然CD 14 mRNA在非拉伸HPLC中不表达，在HPLC中，表达被循环拉伸力强烈诱导4- 8h。在HPLC表面和上清液中分别施加24 h的拉伸力。在拉伸和非拉伸的HPLC上清中均观察到IL-6的产生，但在HPLC上，经拉伸力和中间青霉LPS的连续刺激后，IL-6的产生增加。H7和抗CD 14单克隆抗体MY 4也抑制了IL-14的增强。剪切应力反应元件（SSRE）的共有序列分析表明，剪切应力反应元件（SSRE）的共有序列与剪切应力反应元件（SSRE）的共有序列一致。在CD 14基因的5'侧翼区发现了一个新的突变。另外。结果表明，循环牵张力通过激活PKC诱导细胞表达CD 14，而SSRE与CD 14的表达有关。此外，CD 14的表达可能增加HPLC对LPS的敏感性。少

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