Suzuki K
Department of Orthopaedic Surgery, Hokkaido University School of Medicine, Japan.
Nihon Seikeigeka Gakkai Zasshi. 1990 Oct;64(10):928-36.
A growth factor was purified from human bone matrix by extraction with EDTA, acetone treatment, gel filtration column chromatography, ion exchange column chromatography, and reversed-phase HPLC. Purified protein migrated as a single band to an area with a molecular weight of about 6,000 on SDS-polyacrylamide gel. The purified protein stimulated dose dependent osteoblast proliferation. The sequence of the first 30 N-terminal amino acids of the protein was identical to that of the human insulin-like growth factor-II (IGF-II). MC3T3-E1 cells reacted immunocytochemically to the monoclonal antibody against IGF-II. The culture medium of MC3T3-E1 cells contained immunoreactive IGF-II, which was measured by an enzyme immunoassay. These results indicate that IGF-II is contained in bone matrix, is effective in osteoblast proliferation, produced by the osteoblasts, and secreted from the osteoblasts. Taken together, these results show that IGF-II is present as a local growth factor in the bone system, playing an important role in bone formation following bone resorption.
通过用乙二胺四乙酸（EDTA）提取、丙酮处理、凝胶过滤柱色谱法、离子交换柱色谱法和反相高效液相色谱法，从人骨基质中纯化出一种生长因子。纯化后的蛋白质在十二烷基硫酸钠-聚丙烯酰胺凝胶（SDS-聚丙烯酰胺凝胶）上迁移为一条单一的条带，至分子量约为6000的区域。纯化后的蛋白质刺激成骨细胞增殖呈剂量依赖性。该蛋白质前30个N端氨基酸的序列与人胰岛素样生长因子-II（IGF-II）的序列相同。MC3T3-E1细胞对针对IGF-II的单克隆抗体产生免疫细胞化学反应。MC3T3-E1细胞的培养基中含有免疫反应性IGF-II，通过酶免疫测定法进行测量。这些结果表明，IGF-II存在于骨基质中，对成骨细胞增殖有效，由成骨细胞产生，并从成骨细胞中分泌出来。综上所述，这些结果表明IGF-II作为一种局部生长因子存在于骨系统中，在骨吸收后的骨形成中起重要作用。
