巨噬细胞源成纤维细胞生长因子的分离纯化

为探讨二氧化硅在大鼠肺内引发的一系列纤维增生反应，试图寻找一种关键的细胞增生因子，采用高压液相色谱技术，包括凝胶过滤柱层析、离子交换柱层析以及反相Ｃ４高压液相色谱柱层析，获得一种新的巨噬细胞源成纤维细胞生长因子（ａｌｖｅｏｌａｒｍａｃｒｏｐｈａｇｅ－ｄｅｒｉｖｅｄｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃ－ｔｏｒ，ＡＭＤＧＦ），ＳＤＳ－ＰＡＧＥ结果表明它已达到均一的纯度，其分子量为５８０００，ｐＩ为４．７．该因子的分子量与ＭΦ在体外接受石英粉尘刺激分泌的因子的分子量明显不同．Ｎ端序列测定结果显示它与已知蛋白序列的同源性小于５０％，其刺激成纤维细胞增殖的最适浓度范围为２．０～６．０μｇ／ｍｌ，推测ＡＭＤＧＦ是矽肺纤维化病变发生、发展过程中调节成纤维细胞增殖的重要因子之一

Purification and Characterization of a Macrophage derived Fibroblast Growth Factor

In order to study the fibrogenic reactions in rat lung induced by SiO 2 and to find out the related growth factor,broncho alveolar large and alveolar macrophage were isolated and cultured .A new macrophage derived fibroblast growth factor(AMDGF)which was different from that obtained by in vitro tests was isolated and characterized from both silicotic rat lung lavage macrophages and its culture media.The AMDGF was purified to homogeneous on the basis of sodium dodecyl sulfate polyacrylamide gel electrophoresis by a combination of gel permeation,ion exchange and reverse phase high performance liquid chromatography.The final purified AMDGF had a molecular weight of 58 000(reduced)and a p I of 4 7.AMDGF improvd the proliferation activity of fibroblast at optimum concentration range of 2 0～6 0 μg/ml in culture medium.N terminal amino sequences analysis of AMDGF showed that this protein was of less than 50% homogeneity with all known proteins.It is considered that this factor is one of the main mediators that regulate fibroblast proliferation during the process of silica inducing fibrosis.