Modification of Athferoma by cytokin gene transfer

通过细胞因子基因转移修饰动脉粥样硬化

基本信息

批准号：
09557074
负责人：
SAITO Yasushi
金额：
$ 8.19万
依托单位：
Chiba University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09557074/
关键词：
intima athevosclevosis cytokine re-stenosis ECM 平滑筋細胞遺伝子発現

项目摘要

We developed a rat double-balloon injury model and studied the thickened neointima using immunohistochemical and RT-PCR methods. Fourteen days after the first balloon injury of the rat left common carotid artery, a second balloon injury was inflicted in the same place. Histochemical sections taken 14 days after the single- and double-balloon injuries were used for calculating intimal/medial (I/M) area ratios, as an indicator of neointimal formation, and were subjected to immunohistochemical staining. Total RNA was also purified from some arteries and mRNA expression of some extracellular matrices (ECM) and cytokine receptors related to ECM metabolism was estimated using the RT-PCR method. The I/M ratio in the rat double-balloon injury model (II) (1.84±0.62(mean±SE, n=5)) was significantly (p<0.05) higher than that in the single-balloon injury model (I) (1.30±0.19, n=10). The cell number per neointimal area was less in II than in I. As for the phenotype of smooth muscle cells, α-actin staining showed that the neointima of II consisted of more contractile form than synthetic, whereas that of I consisted of more synthetic form than contractile. The neointima of II was strongly stained with laminin and fibronectin, but that of I was stained only weakly. Consistent with these data, laminin and fibronectin mRNAs were markedly expressed in the neointima of II. Neointimas of both I and II were also stained positively with PDGF (α and β) and TGF-β (types I and II) receptors to the same extent. These results show: that ECM accumulation, particularly of laminin and fibronectin, characterizes the double-balloon injury model; that the marked accumulation of ECM in this model is due to a mechanism other than the PDGF or TGF-β signalling pathway; and that this model resembles the lesion of post-PTCA re-stenosis, and therefore provides the key for its investigation.

我们开发了一种大鼠双球损伤模型，并使用免疫组织化学和RT-PCR方法研究了增厚的新内膜。大鼠第一次气球受伤后十四天后，在同一位置造成了第二次气球损伤。组织化学切片在单球损伤后14天用于计算内膜/培养基（I/M）面积比，作为新内膜形成的指标，并进行免疫组织化学染色。还使用RT-PCR方法估算了一些细胞外胎盘（ECM）（ECM）（ECM）（ECM）（ECM）（ECM）和与ECM代谢相关的细胞因子受体的mRNA表达纯化的总RNA。大鼠双球损伤模型（II）（1.84±0.62（平均±SE，n = 5））中的I/M比显着（P <0.05），比单球损伤模型（I）（I）（1.30±0.19，n = 10）高（p <0.05）。在II中，每个新内膜区域的细胞数量小于I。对于平滑肌细胞的表型，α-肌动蛋白的染色表明，II的新内膜由比合成的更收缩的形式组成，而I的合成形式比收缩更合成形式。 II的新内膜用层粘连蛋白和纤连蛋白强烈染色，但我的粘液蛋白仅被微弱染色。与这些数据一致，层粘连蛋白和纤连蛋白mRNA在II的新内膜中显着表达。 I和II的新内膜也用PDGF（α和β）和TGF-β（I型和II）受体呈阳性染色。这些结果表明：ECM积累，尤其是层粘连蛋白和纤连蛋白的积累，是双球损伤模型的特征。 ECM在该模型中的明显积累是由于PDGF或TGF-β信号通路以外的机制。并且该模型类似于PTCA后重新发生的病变，因此为其投资提供了关键。