Borromeo V, Bramani S, Holder A T, Carter C, Secchi C, Beattie J
Institute of Veterinary Physiology, University of Milan, Italy.
Mol Cell Biochem. 1996 Sep 20;162(2):145-51. doi: 10.1007/BF00227542.
Using monolayer cultures of costal chondrocytes established from four week old Clun Forest lambs, we have demonstrated that, under serum free conditions the cells release three IGFBPs (32, 29 and 21 kDa) into the medium. The most abundant of these--the 32 kDa BP--was shown to be IGFBP-2 by Western blotting. Furthermore we demonstrate that the levels of IGFBP-2 in conditioned medium are acutely increased (6, 12 and 24 h time points) following treatment of cells with bovine GH (1-100 ng/ml). In a parallel set of experiments, using ovine fibroblasts (derived from dermis) we show that IGFBPs of Mr 32, 29 and 21 kDa are also secreted by this cell type. However the relative abundance of these BPs differed from that seen in the chondrocyte cultures, with the 21 kDa species now the most abundant. In addition, prolonged exposure of autoradiographs indicated that fibroblasts secreted a higher Mr IGFBP (most probably ovine BP-3) that was not detected in any of our chondrocyte cultures. Most significant however was the demonstration that bGH did not dramatically affect the levels of IGFBPs in fibroblast cell cultures. We conclude that GH stimulates BP-2 production from chondrocytes and this is a cell-type specific effect in as much as it is not replicated in cultures of dermal fibroblasts.
利用从4周龄克伦森林羔羊建立的肋软骨细胞单层培养物,我们已经证明,在无血清条件下,细胞向培养基中释放三种胰岛素样生长因子结合蛋白(IGFBPs,分子量分别为32、29和21 kDa)。其中最丰富的——32 kDa结合蛋白——经蛋白质免疫印迹法显示为IGFBP - 2。此外,我们还证明,在用牛生长激素(1 - 100 ng/ml)处理细胞后,条件培养基中IGFBP - 2的水平在6、12和24小时时间点急剧增加。在一组平行实验中,使用绵羊成纤维细胞(来源于真皮),我们发现该细胞类型也分泌分子量为32、29和21 kDa的IGFBPs。然而,这些结合蛋白的相对丰度与软骨细胞培养物中的不同,现在21 kDa的种类最为丰富。此外,放射自显影片的长时间曝光表明,成纤维细胞分泌一种分子量更高的IGFBP(很可能是绵羊BP - 3),而在我们的任何软骨细胞培养物中都未检测到。然而,最显著的是,证明牛生长激素并未显著影响成纤维细胞培养物中IGFBPs的水平。我们得出结论,生长激素刺激软骨细胞产生BP - 2,这是一种细胞类型特异性效应,因为在真皮成纤维细胞培养物中未观察到这种效应的重现。