The osteocalcin is a marker of bone metabolism on periodontal disease during of bone resorption

骨钙素是牙周病骨吸收过程中骨代谢的标志物

• 批准号: 05807182
• 负责人: KURIHARA Noriyoshi
• 金额: $ 1.09万
• 依托单位: Meikai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05807182/
• 关键词: osteocalcin; osteoblast; osteoclast; 骨吸収; 歯周炎

A specific immunoassay system for N-terminal osteocalcin (N-OC) was developed to determine if osteocalcin molecules are released from bone degraded by osteoclasts in vitro as a novel method for measuring bone resorption. The assay system employs a monoclonal antibody to the first 20 N-terminal residues of osteocalcin as the solid phase and polyclonal antibodies against these same residues as an enzyme conjugate. This assay system could also detect N-terminal portion of osteocalcin, formed during the degradation of the human osteocalcin molecule by trypsin or cathepsin D.We isolated osteoclasts from human alveolar bone, and cultured them on human bone slices. Using this N-OC assay method, we measured the osteocalcin content in the media from these cultures. Predominantly N-terminal osteocalcin increased in the culture medium, along with a slight increase in the level of intact osteocalcin, during osteoclastic bone resorption. In the presence of IL-1b and IL-6, stimulators of osteoclastic bone resorption, the level of N-terminal osteocalcin increased by 1.5 fold compared with that in the absence of the stimulators. On the contrary, in the presence of bisphosphonate (YM175) or a cathepsin D inhibitor (E-64), inhibitors of osteoclastic bone resorption, the release of N-terminal osteocalcin decreased. These results suggests that the fragments of osteocalcin released during osteoclastic bone resorption and the detection of N-terminal osteocalcin may provide a useful index for bone resorption in vitro.

建立了一种N-末端骨钙素（N-OC）的特异性免疫分析系统，用于检测破骨细胞体外降解骨中骨钙素分子的释放，作为一种新的骨吸收检测方法。该测定系统采用针对骨钙素的前20个N-末端残基的单克隆抗体作为固相，并采用针对这些相同残基的多克隆抗体作为酶缀合物。该检测系统也可以检测N-末端部分的骨钙蛋白，形成的人骨钙蛋白分子的降解过程中，胰蛋白酶或组织蛋白酶D。我们分离破骨细胞从人牙槽骨，并培养他们的人骨切片。使用这种N-OC测定方法，我们测量了来自这些培养物的培养基中的骨钙素含量。主要是N-末端骨钙素增加的培养基中，沿着与完整的骨钙素的水平略有增加，在骨细胞骨吸收。在IL-1b和IL-6的存在下，骨钙素的N-末端的水平增加了1.5倍，相比，在没有刺激剂的情况下。相反，在双膦酸盐（YM 175）或组织蛋白酶D抑制剂（E-64）的存在下，骨钙素骨吸收的抑制剂，N-末端骨钙素的释放减少。结果提示，骨钙素在骨吸收过程中释放的片段及N端骨钙素的检测可为骨吸收的体外研究提供一个有用的指标。