Maciag T, Cerundolo J, Ilsley S, Kelley P R, Forand R
Proc Natl Acad Sci U S A. 1979 Nov;76(11):5674-8. doi: 10.1073/pnas.76.11.5674.
Extracts of bovine hypothalamus were found to contain a significant level of mitogenic activity when tested in a Swiss 3T3 cell [3H]dThd incorporation assay and in a human umbilical vein endothelial cell growth assay. The mitogenic activity responsible for 3T3 cell activity was purified and characterized as a fibroblast growth factor (FGF)-like mitogen. Neither the biologically active FGF-like mitogen purified from the hypothalamus extracts nor FGF purified from bovine pituitary glands was mitogenic when added to human endothelial cells in vitro, suggesting the presence of more than one mitogen in the hypothalamic extracts. The 3T3 and endothelial cell biological activities of hypothalamic extracts were both found to be inactivated by trypsin, subtilisin, and heat treatment, but were stable to dialysis. The endothelial cell growth factor activity could be efficiently separated from the FGF activity by gel exclusion chromatography. The endothelial cell mitogen possessed a molecular weight of approximately 75,000, whereas that of FGF was approximately 15,000. The endothelial cell growth factor activity was found to be inactivated with reducing agents whereas the 3T3 cell mitogenic activity was stable after incubation with 2-mercaptoethanol. Significant levels of endothelial cell mitogenic activity were also found in extracts of bovine brain and pituitary glands.
在瑞士3T3细胞[³H]dThd掺入试验和人脐静脉内皮细胞生长试验中检测时,发现牛下丘脑提取物含有显著水平的促有丝分裂活性。负责3T3细胞活性的促有丝分裂活性被纯化并鉴定为成纤维细胞生长因子(FGF)样促有丝分裂原。从下丘脑提取物中纯化的具有生物活性的FGF样促有丝分裂原和从牛垂体中纯化的FGF,在体外添加到人内皮细胞时均无促有丝分裂作用,这表明下丘脑提取物中存在不止一种促有丝分裂原。下丘脑提取物的3T3细胞和内皮细胞生物活性均被胰蛋白酶、枯草杆菌蛋白酶和热处理灭活,但对透析稳定。通过凝胶排阻色谱法可将内皮细胞生长因子活性与FGF活性有效分离。内皮细胞促有丝分裂原的分子量约为75,000,而FGF的分子量约为15,000。发现内皮细胞生长因子活性被还原剂灭活,而3T3细胞促有丝分裂活性在与2-巯基乙醇孵育后稳定。在牛脑和垂体提取物中也发现了显著水平的内皮细胞促有丝分裂活性。