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Development of drug deliumy system for targeting to injured myocardium by a novel glycoside-binding protein

开发一种新型糖苷结合蛋白靶向损伤心肌的药物谵妄系统

基本信息

批准号：
18590764
负责人：
ISE Hirohiko
金额：
$ 2.43万
依托单位：
Shinshu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590764/
关键词：
Drug deliverv system lectin vimentin desmin GlcNAc 糖鎖心筋細胞血管平滑筋細胞

项目摘要

We investigated a novel lectin that cardiomyocytes and vascular smooth muscle cells (VSMCs) express on cell surface, in order to elucidate the role of carbohydrate-modifications for regeneration and inflammation in cardiovascular diseases and to develop a drug delivery system by using a lectin. We examined the interaction of various glycoside chains with cardiomyocytes and VSMCs and cardiomyocytes by using glycoside-binding assay with various glycoside-bearing polymers-coated dishes. Cardiomyocytes and VSMCs had the interaction with N-acetylglucosamine (GIcNAc) specifically. Moreover, cardiomyocytes and VSMCs took up the GIcNAc-conjugated liposomes. From these results, we assumed that these cells have a GIcNAc-binding lectin on cell surface and tried to identify the lectin. A GIcNAc-recognizing lectin was identified as desmin and vimentin in cardiomyocytes and as vimentin in VSMCs by using two-dimensional electrophoresis and western blotting with GIcNAc-bearing polymer. To confirm whether vimentin and desmin bind to GIcNAc, we examined a binding capacity of vimentin and desmin for GIcNAc by using BIACORE. Vimentin and desmin interacted with GIcNAc-bearing polymer highly and specifically, whereas not glucose- and galactose-bearing polymers. Moreover, to examine whether these protein express on cell surface. we performed double-immunostaining of these cells with anti-vimentin or desmin antibody and fluorescence-conjugated GIcNAc-bearing polymer. We observed that these antibodies and polymer stained on the same region of cell surface by using laser scanning confocal microscopy. From these results, we suggested that as a novel function of vimentin and desmin. these proteins have a binding capacity for GIcNAc and express on surface of cardiomyocytes and vascular smooth muscle cells.

我们研究了心肌细胞和血管平滑肌细胞（VSMCs）在细胞表面表达的一种新型凝集素，以阐明碳水化合物修饰在心血管疾病中的再生和炎症中的作用，并通过使用凝集素开发药物递送系统。我们用不同的糖苷聚合物包被的培养皿，通过糖苷结合试验检测了不同的糖苷链与心肌细胞和VSMCs以及心肌细胞的相互作用。心肌细胞和VSMCs与N-乙酰氨基葡萄糖（GlcNAc）特异性相互作用。此外，心肌细胞和VSMC摄取了GIcNAc缀合的脂质体。根据这些结果，我们假设这些细胞在细胞表面上具有GlcNAc结合凝集素，并试图鉴定凝集素。通过双向电泳和免疫印迹技术，在心肌细胞和血管平滑肌细胞中分别鉴定出GIcNAc识别的凝集素为结蛋白和波形蛋白。为了确认波形蛋白和结蛋白是否与GIcNAc结合，我们通过使用BIACORE检查了波形蛋白和结蛋白对GIcNAc的结合能力。波形蛋白和结蛋白相互作用的GlcNAc轴承聚合物高度和具体的，而不是葡萄糖和半乳糖轴承聚合物。并检测这些蛋白质是否在细胞表面表达。我们用抗波形蛋白或结蛋白抗体和荧光缀合的携带GlcNAc的聚合物对这些细胞进行双重免疫染色。激光扫描共聚焦显微镜观察到这些抗体和聚合物在细胞表面的同一区域染色。从这些结果中，我们建议作为一个新的功能的波形蛋白和结蛋白。这些蛋白质具有与GlcNAc结合的能力，并在心肌细胞和血管平滑肌细胞的表面上表达。