摘要
目的探讨周期性张应变对人牙周膜细胞(human periodontal ligament cells,hPDLCs)增殖的影响及其相关机制。方法应用FX-4000T细胞应变加载系统,对体外培养的hPDLCs施加周期性张应变,幅度分别为10%和20%,加载时间为6h和24h,频率均为0.1Hz,以未加载的静态细胞作为对照组。应用流式细胞术检测人牙周膜细胞细胞周期的变化,并应用Western blot法检测细胞增殖细胞核抗原(PCNA)和细胞内信号转导分子p-ERK1/2表达的变化。用ERK1/2的特异性抑制剂PD98059预处理细胞后,在0.1Hz,10%幅度条件下,加载6h,检测p-ERK1/2的表达水平变化对细胞PCNA表达的影响。结果与静态组相比,周期性张应变增加了S期人牙周膜细胞的比例,并诱导人牙周膜细胞的PCNA和p-ERK1/2表达增加,10%幅度和20%幅度相比无显著性差异。同一幅度张应变,加载6h和24h对细胞PCNA和p-ERK1/2表达的影响无显著性差异。ERK1/2的抑制剂PD98059不仅可以明显抑制张应变诱导的p-ERK1/2活化,而且张应变诱导的细胞PCNA表达也被明显抑制。结论周期性张应变可激活ERK信号通路,促进体外培养人牙周膜细胞的增殖。
Objective To explore the role of extracellular signal-regulated kinase （ERK） signaling pathway on cyclic strain induced proliferation of human periodontal ligament cells （hPDLCs）. Method The hPDLCs cultured in vitro were subjected to cyclic strain by FX-4000T system with 10 % or 20 %-elongation magnitude, 6 or 24 hours-duration respectively, at the same frequency of 0.1 Hz. The cell cycles of hPDLCs were measured by FCM. The expressions of p-ERK1/2 and PCNA in hPDLCs were analyzed by Western blotting. To investigate the effect of p-ERK on cell proliferation, hPDLCs were incubated with PD98059, a specific ERK kinase inhibitor, and then the expression of PCNAwas also analyzed byWestern blot after hPDLCs were exposed to 10 %-cyclic strain at 0.1 Hz-frequency for 6 hours. Result Cyclic strain obviously increased the ratio of S period in the cell cycles, the expressions of PCNAand p-ERK1/2 in hPDLCs, in which the magnitude and duration of cyclic strain had no significant difference. PD98059 could repress not only the activation of p-ERK1/2 but the expression of PCNA induced by cyclic strain in hPDLCs. Conclusions Cyclic strain promotes the proliferation of hPDLCs through ERK signaling pathway.
出处
《医用生物力学》
CAS
CSCD
2009年第3期211-215,222,共6页
Journal of Medical Biomechanics
基金
国家自然科学基金资助项目(10502034)
关键词
周期性张应变
牙周膜细胞
增殖
增殖细胞核抗原
ERK信号通路
Cyclic strain
Periodontal ligament cells（PDLCs）
Proliferation
Proliferating cell nuclear antigen （PCNA）
ERK signaling pathway
