牵张应力对人牙周膜细胞MMP-13/TIMP-1表达的影响及其信号转导途径研究
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国家自然科学基金资助项目(30300393,30970697)


Expression of MMP-13/TIMP-1 and signal transduction pathways in response to mechanical strain in human periodontal ligament cells
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    摘要:

    目的 研究不同强度机械牵张应力对人牙周膜细胞基质金属蛋白酶-13(MMP-13)及其组织抑制因子-1(TIMP-1)表达的影响,并探讨机械牵张应力作用下人牙周膜细胞MMP-13/TIMP-1表达变化的信号转导途径。方法通过细胞牵张应力加载系统对体外培养的人牙周膜细胞同时施加0%、6%、12%和18%形变率的机械牵张应力,作用24 h后,用RT-PCR方法检测细胞受力后MMP-13/TIMP-1 mRNA表达的变化,用免疫印迹法检测其蛋白表达的变化。另外,通过使用不同信号途径的特异性抑制剂,用RT-PCR方法分别检测不同抑制剂对牵张应力作用下牙周膜细胞MMP-13/TIMP-1 mRNA表达的变化。结果人牙周膜细胞受力后,MMP-13 /TIMP-1 mRNA及蛋白表达随牵张应力强度的增大明显增加。PD098059可抑制机械牵张应力作用下牙周膜细胞MMP-13 mRNA表达的增加。放线菌酮可抑制机械牵张应力作用下TIMP-1 mRNA表达的增加。结论不同强度机械牵张应力可以影响人牙周膜细胞MMP-13/TIMP-1的表达,进而影响牙周组织细胞外基质代谢。机械牵张应力作用下MMP-13表达的增加是通过ERK-MAPK途径。机械牵张应力作用下TIMP-1表达的增加是通过新生蛋白途径。

    Abstract:

    Objective To investigate the effects of different magnitudes of mechanical strain on the expression of MMP-13/TIMP-1 and try to determine the signal transduction pathways in response to mechanical strain in human periodontal ligament cells (HPDLCs) in vitro. MethodHPDLCs were subjected to 0%,6%,12% or 18% elongation for 24 h by using cell stress loading system simultaneously. Then the MMP-13/TIMP-1 mRNA and protein expression in cells were tested by reverse transcription polymerase chain reaction (RT-PCR) and western blotting respectively. Furthermore, specific inhibitors were employed to examine the role of different signal transduction pathway on the expression of straininduced MMP-13 /TIMP-1 in HPDLCs. ResultsThe expression of MMP-13 /TIMP-1 in HPDLCs significantly increased in groups of 6%,12%, 18% elongation in a magnitude-dependent manner compared with the control group (0%) after the mechanical strain treatment for 24 h. PD098059 and cycloheximide could inhibit the increase in MMP-13 and TIMP-1 mRNA expression in response to mechanical strain respectively. Conclusions Different magnitudes of mechanical strain can affect the expression of MMP-13/TIMP-1 in HPDLCs in a magnitudedependent manner, and further affect the periodontium remodeling with the characteristics of degradation and synthesis of extracellular matrix in response to mechanical strain. The ERK-MAPK pathway is involved in straininduced MMP-13 expression while the strain-induced TIMP-1 expression depends on de novo protein synthesis.

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李永明,唐林,张晓东,丁寅.牵张应力对人牙周膜细胞MMP-13/TIMP-1表达的影响及其信号转导途径研究[J].医用生物力学,2010,25(6):422-427

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  • 收稿日期:2010-11-06
  • 最后修改日期:2010-11-22
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