权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Angiogenesis promotion by fixing biotinylated VFGF on endothelial cells

通过将生物素化的 VFGF 固定在内皮细胞上促进血管生成

基本信息

批准号：
13671459
负责人：
HOYA Katsumi
金额：
$ 1.41万
依托单位：
Dokkyo University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671459/
关键词：
avidin endothelium biotin VFGF drug delivery VEGF

项目摘要

We have developed an intravascular drug delivery system, in. which a drug injected from a catheter is fixed to the vasculature of the targeted tissue. Cellular protein of viable endothelial cells is first biotinylated directly by biotinylation reagents, and then bound by an avidinated drug or, using avidin as a linker, a biotinylated drug. We hypothesized that if vascular endothelial growth factor (VEGF), which promotes endothelial proliferation and angiogenesis, is biotinylated and given to the endothelium by the above method, its effect will become greater than when the cells are simply exposed to it for very few minutes. In the first experiment, we investigated intracellular locations of the molecule given to the endothelium by the above method. Streptavidht labeled with β-galactosidase was fixed to biotmylated endothelium, and.cells were harvested 0,1,2,3,6,12 hours after. The enzyme was visualized byhistochemical method (x-gal). Electron microscopic study revealed that the enzyme was located at 0 hour in the cytoplasm and reached nuclear membrane. Few were found on the cellular membrane. In the second experiment, biotinylated VEGF was made, and fixed to the biotinylated endothelial cells. Though the activity of the biotinylated VEGF was the same as that of intact VEGF, the fixation of the growth factor on the cells was not so effective for the proliferarion of cultured endothelial cells. We speculate that drugs that influence via the cell membrane receptors are not good candidates for our drug delivery method because the biotinylated drugs enter too fast into the cells without binding to their receptors.

我们已经开发了一种血管内给药系统。其中从导管注射的药物被固定到目标组织的血管系统。活内皮细胞的细胞蛋白首先被生物素化试剂直接生物素化，然后被抗生物素化药物或使用抗生物素蛋白作为接头的生物素化药物结合。我们假设，如果将促进内皮细胞增殖和血管生成的血管内皮生长因子（VEGF）生物素化并通过上述方法给予内皮细胞，则其效果将比细胞仅暴露于其几分钟时更大。在第一个实验中，我们研究了通过上述方法给予内皮的分子的细胞内位置。将β-半乳糖苷酶标记的Streptavidht固定于生物膜化的内皮细胞上，分别于0、1、2、3、6、12小时收获细胞。用组织化学方法（x-gal）观察酶的表达。电镜观察表明，酶在0小时定位于细胞质，并到达核膜。在细胞膜上发现很少。在第二个实验中，制备生物素化的VEGF，并固定到生物素化的内皮细胞上。虽然生物素标记的VEGF活性与完整的VEGF相同，但生长因子对细胞的固定对培养的内皮细胞的增殖并不那么有效。我们推测，通过细胞膜受体产生影响的药物不是我们药物递送方法的良好候选者，因为生物素化药物进入细胞太快而不结合其受体。