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.

我们建立了大鼠双球囊损伤模型，并使用免疫组织化学和 RT-PCR 方法研究了大鼠左颈总动脉第一次球囊损伤后的十四天，在同一位置进行了第二次球囊损伤。使用单球囊和双球囊损伤后 14 天计算内膜/内侧 (I/M) 面积比，作为新内膜形成的指标，并进行免疫组织化学分析还从一些动脉中纯化了总 RNA，并使用 RT-PCR 方法估计了一些细胞外基质 (ECM) 和与 ECM 代谢相关的细胞因子受体的 mRNA 表达。 II) (1.84±0.62(mean±SE, n=5)) 显着 (p<0.05) 高于单球囊损伤模型 (I) (1.30±0.19, n=10)。II 中每个新内膜区域的细胞数量少于 I。至于平滑肌细胞的表型，α-肌动蛋白染色显示 II 的新内膜比合成的新内膜由更多的收缩形式组成，而 I 的新内膜由更多的收缩形式组成。 II 的新内膜被层粘连蛋白和纤连蛋白强烈染色，但 I 的新内膜仅被弱染色，与这些数据一致，层粘连蛋白和纤连蛋白 mRNA 被显着染色。 I 和 II 的新内膜中表达的 I 和 II 的新内膜也以相同程度被 PDGF（α 和 β）和 TGF-β（I 型和 II 型）受体染色。层粘连蛋白和纤连蛋白，表征了双球囊损伤模型；该模型中 ECM 的显着积累是由于 PDGF 或 TGF-β 信号通路以外的机制造成的，并且该模型类似于PTCA 术后再狭窄，因此为其研究提供了关键。