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Studies on molecular biology of phenotype change of smooth muscle cells

平滑肌细胞表型变化的分子生物学研究

基本信息

批准号：
03670311
负责人：
MORISAKI Nobuhiro
金额：
$ 1.34万
依托单位：
School of Medicine, 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：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

项目摘要

1. Modulation of phenotype of smooth muscle cells: Cultured endothelial cells and macrophages modulated cultured rabbit aortic smooth muscle cells(SMC) through secretion of platelet-derived growth factor(PDGF). Of PDGF isomers, PDGF -BB and -AB but not -AA were the phenotype modulation factors. By these phenotype modulation SMC acquired the following characteristics; 1. rapid growth. 2. expression of scavenger receptors. 3. secretion of SMC derived growth factor(S DGF).2. Phenotype modulation by a ballooning model: Cultured SMC from rabbit aorta treated with a balloon catheter were examined for their phenotype change as a function of time. One day after ballooning SMC phenotype was not changed. But 3 days after SMC growth became rapid. 7 days after SMC showed all characteristics mentioned above. At this time point no intimal layer was formed, indicating that phenotype change occurs in the media.3. Expression of the scavenger receptors in SMC: Scavenger receptors were induced in SMC by treatment with phorbol ester. The mechanism of induction was through release of lysolecithin by activation of phospholipase A2 but not through activation of protein kinase C.4. Characterization of SDGF: The growth activity of SDGF was inhibited by a polyclonal antibody to basic FGF. But western blot analysis showed that the molecular size of SDGF was 32kd but not 17kd, suggesting that SDGF is not FGF itself. Northern blot analysis showed that only 6.4kb band of mRNA OF basic FGF was detected by using cDNA probe for basic FGF gene. These results suggested that SDGF is different from basic FGF. Further cloning is in progressusing the antibody with cross-reactivity with SDGF.

1.平滑肌细胞表型的调节：培养的内皮细胞和巨噬细胞通过分泌血小板衍生生长因子（PDGF）调节培养的兔主动脉平滑肌细胞（SMC）。在PDGF异构体中，PDGF-BB和-AB是表型调节因子，而-AA不是.通过这些表型调控，SMC获得了以下特征：1.快速增长。2.清道夫受体的表达。3. SMC衍生生长因子（SDGF）的分泌。通过球囊模型的表型调节：检查来自用球囊导管处理的兔主动脉的培养SMC的表型变化作为时间的函数。球囊扩张后一天，SMC表型没有改变。但3天后SMC生长迅速。SMC培养7d后，上述特征均出现。此时无内膜层形成，表明中膜发生表型改变. SMC中清道夫受体的表达：用佛波酯处理SMC可诱导清道夫受体。诱导机制是通过激活磷脂酶A2释放溶血素，而不是通过激活蛋白激酶C。SDGF的表征：SDGF的生长活性被碱性FGF的多克隆抗体抑制。但Western blot分析显示SDGF分子大小为32kd而非17kd，提示SDGF本身不是FGF。北方印迹分析表明，碱性FGF基因cDNA探针仅检测到6.4kb的mRNA条带。这些结果表明SDGF不同于碱性FGF。进一步的克隆正在使用与SDGF交叉反应的抗体进行。