骨膜蛋白通过αvβ3 整合素/ERK 通路调控人牙周膜细胞中 MMP-2 的表达。
PERIOSTIN regulates MMP-2 expression via the αvβ3 integrin/ERK pathway in human periodontal ligament cells.
机构信息
Department of Orthodontics and Dentofacial Orthopedics, Institute of Health Biosciences, The University of Tokushima Graduate School, 3-18-15 Kuramoto-cho, Tokushima City, Tokushima, Japan.
出版信息
Arch Oral Biol. 2012 Jan;57(1):52-9. doi: 10.1016/j.archoralbio.2011.07.010. Epub 2011 Aug 31.
OBJECTIVE
During orthodontic tooth movement, activation of the vascular system in the compressed periodontal ligament (PDL), which becomes hypoxic, is essential for periodontal tissue remodelling. PERIOSTIN, an extracellular matrix protein, is expressed in PDL and its concentration is increased on the compressive side during orthodontic tooth movement. PERIOSTIN promotes angiogenesis through upregulation of matrix metalloproteinase (MMP)-2, which has been shown to be expressed via αvβ3 integrin/extracellular signal-related kinase (ERK) signalling pathway and vascular endothelial growth factor (VEGF). Therefore, we hypothesized that hypoxia-induced PERIOSTIN promotes MMP-2 expression via αvβ3 integrin/ERK signalling and VEGF in PDL cells.
METHODS
Human PDL cells were cultured in condition medium containing desferrioxamine (DFO) to mimic hypoxia. The total RNA, cell lysates or supernatant were collected, and MMP2 and VEGF expression, PERIOSTIN expression and ERK phosphorylation, and MMP-2 activity were analysed by real-time RT-PCR, western blot analysis, and zymography, respectively. A recombinant human PERIOSTIN or PERIOSTIN siRNA was applied to the cells, then the total RNA was extracted to measure MMP-2 and VEGF expression. The cells were treated with αvβ3 integrin-blocking antibody or ERK inhibitor followed by PERIOSTIN stimulation. MMP-2 expression was measured by real-time RT-PCR.
RESULTS
PERIOSTIN was upregulated in a time-dependent manner in human PDL cells treated with DFO, a chemical hypoxia mimic. MMP-2 and VEGF expression, and MMP-2 activity were increased by DFO or PERIOSTIN treatment, and decreased by PERIOSTIN silencing. PERIOSTIN treatment also induced ERK phosphorylation, and PERIOSTIN-induced MMP-2 was reduced by αvβ3 integrin-blocking antibody or ERK inhibitor.
CONCLUSION
These data suggest that PERIOSTIN upregulates MMP-2 expression via the αvβ3 integrin/ERK signalling pathway and VEGF expression in human PDL cells.
目的
在正畸牙齿移动过程中，受压牙周韧带（PDL）中血管系统的激活对于牙周组织重塑至关重要，受压牙周韧带会缺氧。骨膜蛋白（PERIOSTIN）是一种细胞外基质蛋白，在 PDL 中表达，在正畸牙齿移动过程中，其浓度在受压侧增加。PERIOSTIN 通过上调基质金属蛋白酶（MMP）-2 促进血管生成，已证明 MMP-2 通过 αvβ3 整联蛋白/细胞外信号调节激酶（ERK）信号通路和血管内皮生长因子（VEGF）表达。因此，我们假设缺氧诱导的 PERIOSTIN 通过 PDL 细胞中的 αvβ3 整联蛋白/ERK 信号和 VEGF 促进 MMP-2 的表达。
方法
将人牙周膜细胞在含有去铁胺（DFO）的条件培养基中培养，以模拟缺氧。收集总 RNA、细胞裂解物或上清液，通过实时 RT-PCR、western blot 分析和明胶酶谱分析分别分析 MMP2 和 VEGF 表达、PERIOSTIN 表达和 ERK 磷酸化以及 MMP-2 活性。应用重组人 PERIOSTIN 或 PERIOSTIN siRNA 处理细胞，然后提取总 RNA 以测量 MMP-2 和 VEGF 表达。用 αvβ3 整联蛋白阻断抗体或 ERK 抑制剂处理细胞，然后用 PERIOSTIN 刺激，通过实时 RT-PCR 测量 MMP-2 表达。
结果
在用人牙周膜细胞用化学缺氧模拟物 DFO 处理的实验中，PERIOSTIN 呈时间依赖性上调。DFO 或 PERIOSTIN 处理可增加 MMP-2 和 VEGF 的表达和 MMP-2 活性，PERIOSTIN 沉默则降低 MMP-2 和 VEGF 的表达和 MMP-2 活性。PERIOSTIN 处理还诱导 ERK 磷酸化，αvβ3 整联蛋白阻断抗体或 ERK 抑制剂可降低 PERIOSTIN 诱导的 MMP-2。
结论
这些数据表明，PERIOSTIN 通过 αvβ3 整联蛋白/ERK 信号通路和 VEGF 表达上调人牙周膜细胞中 MMP-2 的表达。
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