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Studies on autocrine system for migration factor for smooth muscle cells

平滑肌细胞迁移因子自分泌系统的研究

基本信息

批准号：
01570351
负责人：
MORISAKI Nobuhiro
金额：
$ 1.34万
依托单位：
Chiba 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：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570351/
关键词：
smooth muscle cells migration factor rabbit rat atherosclerosis TGF-beta 血小板由来増殖因子 fibronectin

项目摘要

Following results were obtained using cultured rabbit and rat aortic smooth muscle cells :1. Conditioned media from cultured rabbit and rat aortic smooth muscle cells showed high migration activity for smooth muscle cells. The activity was several times as high as that of purified platelet derived growth factor (PDGF). We named this factor smooth muscle cell derived growth factor (SDGF).2. SDGF was chemotactic but not chemokinetic. It was stable at the treatments such as acidification, alkalization, freezing and thawing, and reduction but was unstable at the treatments such as heating and tripsinization. It was not dializable against the membrane with cutoff pore size 3500. The activity was not neutralized by the antibodies against PDGF or fibronectin. These results suggested that SDMF was a new factor different from known migration factors for smooth muscle cells.3. Secretion of SDMF was 2-3 fold higher in the smooth muscle cells from atherosclerotic intima than that in the smooth muscle cells from normal media.4. Preincubation with transforming growth factor- beta reduced secretion of SDMF from smooth muscle cells.5. Purification : SDMF was adsorbed to heparin-column and eluted by 1M NaC1. The peak of SDMF was collected, dialized, reophilized, and then subjected to a high performance liquid chromatography (HPLC superose 6). Two peaks were obtained at molecular size 400KD and 40KD. The latter peak was again sudjected to HPLC superose 12 with eluate containing SDS. Two peaks were obtained at molecular size 25KD and less than 500D. This purification is now in progress.

采用体外培养的兔和大鼠主动脉平滑肌细胞，获得以下结果：1.从培养的兔和大鼠主动脉平滑肌细胞的条件培养基显示平滑肌细胞的高迁移活性。其活性是纯化血小板衍生生长因子（PDGF）的数倍。我们将此因子命名为平滑肌细胞衍生生长因子（smooth muscle cell derived growth factor，SDGF）. SDGF是趋化性的，但不是趋化动力学的。在酸化、碱化、冻融、还原等条件下稳定，在加热、胰蛋白酶化等条件下不稳定。它对截止孔径为3500的膜不可透析。活性不被针对PDGF或纤连蛋白的抗体中和。这些结果提示SDMF是一种不同于已知平滑肌细胞迁移因子的新因子.动脉粥样硬化内膜平滑肌细胞分泌SDMF的量是正常中膜平滑肌细胞的2-3倍.用转化生长因子-β预孵育可减少平滑肌细胞分泌SDMF。纯化：将SDMF吸附到肝素柱上，并用IM NaCl洗脱。收集SDMF峰，透析，再干燥，然后进行高效液相色谱（HPLC superose 6）。在分子量400 KD和40 KD处有两个峰。将后一峰再次用含有SDS的HPLC superose 12洗脱。在分子量25 KD和小于500 D处有两个峰。这种净化正在进行中。