Role of in glomerular injury

我们之前已经证明了一种抗黏附素分子——足霉素（podocalyxin）与接头蛋白结合，这种相互作用对维持足细胞结构的独特组织非常重要。在本研究项目中，我们重点关注足alyxin糖基化的改变，这可能会影响肾小球粘连，这是严重肾脏疾病的标志。我们研究了哪些糖转移酶在足细胞中特异性表达，并进行了2-D SDS-PAGE检测足霉霉素糖基化的改变。此外，我们还检测了新型基质材料蜂窝状膜是否为足细胞培养提供了合适的周长。我们发现两种独特的糖转移酶（糖蛋白- n-乙酰半乳糖胺-3-13-半乳糖基转移酶，C1Gal-t-1, UDP-G1cNAc 2- epimase， GNE）在体外和体内足细胞中表达。在蜂窝状膜上培养足细胞时，足细胞特异性mRNA （nephrin, CD2AP, podocalyxin）表达增加。扫描电镜观察在该膜上培养的多层细胞。我们还发现足细胞中足alyxin与衔接蛋白（NHERF2和ezrin）的相互作用与血管紧张素II浓度成比例地降低。由于技术问题，我们未能对纯化的足霉霉素进行糖链分析。

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.