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Role of in glomerular injury

在肾小球损伤中的作用

基本信息

批准号：
18590885
负责人：
TAKEDA Tetsuro
金额：
$ 2.5万
依托单位：
Niigata 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

项目摘要

We have previously demonstrated that an anti-adhesin molecule, podocalyxin binds to the adaptor proteins and that the interaction is important to maintain the unique organization of podocyte structure. In this research project, we focused on altered glycation of podocalyxin that should affect glomerular adhesion, indicating a marker of severe kidney diseases. We investigated what kind of sugar transferases are specifically expressed in podocyte and carried out 2-D SDS-PAGE to detect the altered glycation of podocalyxin. Also, we tested podocyte-specific mRNA expression whether novel material of substratum, honeycomb patterned film provides suitable circumference for podocyte culture.1. We found that two unique sugar transferases (glycoprotein-N-acetylgalactosamine-3-13-galactosyltransferase, C1Gal-t-1, UDP-G1cNAc 2-epimerase, GNE) are expressed in podocyte in vitro and in vivo by RT-PCR.2. Podocyte-specific mRNA (nephrin, CD2AP, podocalyxin) expression was increased in podocyte cultured on honeycomb patterned-film. Multi-layered cells cultured on this film were observed by scanning electron microscopy.3. We also found that the interaction between podocalyxin and adaptor proteins (NHERF2 and ezrin) in podocyte cell lines was reduced in proportion to the angiotensin II concentration.4. We failed to analyze sugar chains of purified podocalyxin, because to technical problems.

我们以前已经证明了抗黏附素分子podocalysin与接头蛋白结合，这种相互作用对于维持足细胞结构的独特组织是重要的。在这项研究项目中，我们重点研究了影响肾小球粘连的Podocalyx糖基化改变，这表明它是严重肾脏疾病的标志。我们研究了什么类型的糖转移酶是在足细胞中特异表达的，并进行了二维SDS-PAGE来检测足细胞中足细胞糖基化的改变。此外，我们还检测了新型基质材料蜂窝花纹薄膜是否为足细胞培养提供了合适的周长，并检测了足细胞特异性mRNA的表达。RT-PCR法发现两种独特的糖转移酶(glycoprotein-N-acetylgalactosamine-3-13-galactosyltransferase，C1Gal-t-1，UdP-G1cNAc2-epimerase，GnE在体外和体内足细胞中均有表达。在蜂窝图案膜上培养的足细胞中，足细胞特异性mRNA(newitin、CD2AP、podocalyx)的表达增加。扫描电子显微镜下观察到在该膜上培养的多层细胞。我们还发现，足细胞系中Podocalyx与接头蛋白(NHERF2和Ezrin)之间的相互作用随着血管紧张素II浓度的增加而降低。由于技术上的原因，我们未能对纯化后的香豆素进行糖链分析。