Effect of compressive force on the expression of MMPs, PAs, and their inhibitors in osteoblastic Saos-2 cells
Effect of compressive force on the expression of MMPs, PAs, and their inhibitors in osteoblastic Saos-2 cells
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DOI：
10.1016/j.lfs.2006.01.040
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发表时间：
2006-07-04
期刊：
影响因子：
6.1
通讯作者：
Maeno, Masao
中科院分区：
文献类型：
--
作者：
Mitsui, Narihiro;Suzuki, Naoto;Maeno, Masao
骨基质转换受基质金属蛋白酶（MMPs）、基质金属蛋白酶组织抑制剂（TIMPs）和纤溶酶原激活系统（包括组织型纤溶酶原激活物（tPA）、尿激酶型纤溶酶原激活物（uPA）和纤溶酶原激活物抑制剂1型（派-1））的调节。我们以前证明了1.0g/cm（2）的压力是诱导成骨细胞Saos-2成骨的最佳条件。在这里，我们研究了机械应力对Saos-2细胞中MMPs、TIMPs、tPA、uPA和派-1表达的影响。细胞在含10%胎牛血清的Dulbecco改良Eagle培养基中培养24小时，并在有或无持续压力（0.5-3.0g/cm（2））的情况下培养。通过使用实时PCR测定mRNA水平来估计MMPs、TIMPs、uPA、tPA和派-1基因表达水平，并且使用ELISA测定蛋白水平。当压力为1.0 g/cm 2时，MMP-1、MMP-2、MMP-14和TIMP-1的表达水平明显超过对照组水平，而当压力为3.0 g/cm 2时，MMP-3、MMP-13、TIMP-2、TIMP-3、TIMP-4、tPA、uPA和派-1的表达水平明显超过对照组水平。这些结果表明，机械应力刺激骨基质周转增加这些蛋白酶和抑制剂，骨基质蛋白的蛋白水解降解的机制与机械应力的强度不同。(c)2006年爱思唯尔公司All rights reserved.
Bone matrix turnover is regulated by matrix metalloproteinases (MMPs), tissue inhibitors of matrix metalloproteinases (TIMPs), and the plasminogen activation system, including tissue-type plasminogen activator (tPA), urokinase-type plasminogen activator (uPA), and plasminogen activator inhibitor type-1 (PAI-1). We previously demonstrated that 1.0g/cm(2) of compressive force was an optimal condition for inducing bone formation by ostcoblastic Saos-2 cells. Here, we examined the effect of mechanical stress on the expression of MMPs, TIMPs, tPA, uPA, and PAI-1 in Saos-2 cells. The cells were cultured in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum and with or without continuously compressive force (0.5-3.0g/cm(2)) for up to 24h. The levels of MMPs, TIMPs, uPA, tPA, and PAI-1 gene expression were estimated by determining the mRNA levels using real-time PCR, and the protein levels were determined using ELISA. The expression levels of MMP-1, MMP-2, MMP-14, and TIMP-1 markedly exceeded the control levels at 1.0 g/cm(2) of compressive force, whereas the expression levels of MMP-3, MMP-13, TIMP-2, TIMP-3, TIMP-4, tPA, uPA, and PAI-1 markedly exceeded the control levels at 3.0g/cm(2). These results suggest that mechanical stress stimulates bone matrix turnover by increasing these proteinases and inhibitors, and that the mechanism for the proteolytic degradation of bone matrix proteins differs with the strength of the mechanical stress. (c) 2006 Elsevier Inc. All rights reserved.