Periodontal ligament cells under intermittent tensile stress regulate mRNA expression of osteoprotegerin and tissue inhibitor of matrix metalloprotease-1 and -2
Periodontal ligament cells under intermittent tensile stress regulate mRNA expression of osteoprotegerin and tissue inhibitor of matrix metalloprotease-1 and -2
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DOI：
10.1007/s00774-003-0456-0
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发表时间：
2004-04
影响因子：
3.3
通讯作者：
K. Tsuji;Keiji Uno;Gui-xia Zhang;M. Tamura
中科院分区：
文献类型：
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作者：
K. Tsuji;Keiji Uno;Gui-xia Zhang;M. Tamura
采用Flexercell应变仪研究了人牙周膜（PDL）细胞在间歇性张应力作用下骨保护素（OPG）、核因子-κ B配体受体激活因子（RANKL）、基质金属蛋白酶组织抑制剂（TIMP）-1和-2、基质金属蛋白酶（MMP）-1和-2的mRNA表达。逆转录酶-聚合酶链反应分析表明，机械力上调OPG mRNA。我们还表明，OPG在条件培养基中的蛋白质浓度增加后，负荷与拉伸应力，确定通过酶联免疫吸附测定。TIMP-1和-2 mRNA水平也增加，而RANKL、MMP-1和MMP-2 mRNA水平几乎不受影响。我们进一步研究了张应力负荷和添加流产沙门氏菌平衡多糖（LPS）对PDL细胞mRNA表达的影响。机械应变诱导的OPG mRNA的量被发现减少与添加LPS的文化。在吲哚美辛或染料木素存在下，拉伸诱导的OPG mRNA表达被抑制，而环己酰亚胺的加入抑制拉伸诱导的TIMP-1 mRNA表达，表明张应力通过诱导OPG和TIMP-1 mRNA表达调节PDL细胞中的环氧合酶活性、酪氨酸磷酸化和从头蛋白合成。这些结果提供的证据表明，拉伸的机械刺激是负责观察到的PDL组织中的骨吸收和组织降解的调节。
We studied the mRNA expression of osteoprotegerin (OPG), receptor activator of NF-kappa B ligand (RANKL), tissue inhibitor of matrix metalloprotease (TIMP)-1 and -2, and matrix metalloprotease (MMP)-1 and -2 by human periodontal ligament (PDL) cells under intermittent tensile stress using a Flexercell Strain Unit. Analysis by reverse transcriptase-polymerase chain reaction showed that mechanical force upregulated OPG mRNA. We also demonstrated that the protein concentration of OPG in conditioned medium increased upon loading with tensile stress, as determined by enzyme-linked immunosorbent assay. TIMP-1 and -2 mRNA levels also increased, whereas levels of RANKL, MMP-1, and MMP-2 mRNA were barely affected. We further examined the effect of loading with tensile stress and addition ofSalmonella abortus equilipopolysaccharide (LPS) on the mRNA expression of PDL cells. The amount of OPG mRNA induced by mechanical strain was found to decrease with the addition of LPS to cultures. The induction of OPG mRNA expression by stretching was inhibited in the presence of indomethacin or genistein, whereas TIMP-1 mRNA expression induced by stretching was inhibited by the addition of cycloheximide, suggesting that tensile stress regulates cyclooxygenase activities, tyrosine phosphorylation, and de novo protein synthesis in PDL cells through the induction of OPG and TIMP-1 mRNA expression. These results provide evidence that the mechanical stimulus of stretching is responsible for the observed regulation of bone resorption and tissue degradation in PDL tissue.