ADVANCED GLYCATION ENDPRODUCTS, THEIR RECEPTORS, AND VASCULAR DISEASE

高级糖化终产物、其受体和血管疾病

• 批准号: 6097890
• 负责人: MICHAEL T CROW
• 金额: --
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6097890
• 关键词: Animalia; aging; animal old age; biological signal transduction; cardiovascular disorder; genetically modified animals; glycation; molecular cloning; nuclear factor kappa beta; oxidative stress; protein metabolism; receptor; receptor expression; transfection; vascular endothelium; vascular smooth muscle

SUMMARY OF WORK Advanced glycation endproducts of proteins (AGE) accumulate in the plasma and in tissues with advancing age and at an accelerated rate in diabetes. Our previous work has shown that AGEs induce a pro-oxidant stress in vascular cells, leading to increased monocyte chemoattractant protein-1(MCP-1) production and increased PDGF B chain activity, both of which have been implicated in vascular lesion development. Our current research efforts are focussed on identifying the signaling pathways by which AGEs affect gene expression so that molecular strategies can be developed to block their effects. Our results indicate that signaling is initiated by the binding of AGEs to a unique receptor called RAGE (Receptor for AGEs). We have cloned RAGE from rat intimal vascular smooth muscle cells and constructed epitope-tagged wild type and mutant receptors and shown that transfection of wild type receptor leads to increased MCP-1 RNA levels in response to AGEs. Mutant receptors in which the cytosolic tail has been removed, however, fail to signal increased MCP-1 production in response to AGE stimulation, and, in fact, block the ability of either endogenous or transfected wild type receptors to signal. Two hybrid screening in yeast have identified a number of proteins that interact with the cytosolic tail of RAGE. These include the adapter protein, shc, which has been implicated in receptor mediated activation of MAPKinase pathways, a previously identified protein of unknown function known as p37NB, and several enzymes associated with protein ubiquination, which have recently been shown by others to associate with other signaling receptors only in response to ligand activation. These observations demonstrate that the cytosolic tail of RAGE can engage intracellular proteins important in signal transduction that may be responsible for AGE-induced changes in gene expression.

晚期糖化 蛋白质终产物（AGE）在血浆中积累， 组织随着年龄的增长和糖尿病的加速。 我们以前的工作表明AGEs诱导一种促氧化剂 血管细胞应激，导致单核细胞增多 趋化蛋白-1（MCP-1）的产生和PDGF的增加 B链活性，这两者都与血管 病变发展我们目前的研究工作集中在 确定AGEs影响基因表达的信号通路 表达，以便可以开发分子策略来阻断 他们的影响。我们的研究结果表明，信号是由 AGEs与一种独特的受体结合，称为AGEs（AGEs受体）。 AGEs）。我们从大鼠血管内膜平滑肌中克隆了p53 肌肉细胞和构建的表位标记的野生型和突变体 受体，并显示野生型受体的转染导致 增加MCP-1 RNA水平。突变体 其中胞质尾部已被去除的受体，然而， 未能发出响应AGE的MCP-1产生增加的信号 刺激，事实上，阻止内源性或 转染野生型受体的信号。双杂交筛选法 酵母已经鉴定出许多蛋白质， 胞质尾这些包括衔接蛋白，shc， 它与受体介导的 MAP激酶通路，一种先前鉴定的未知蛋白质 功能称为p37 NB，和几种酶相关 蛋白质泛素化，最近已经被其他人证明， 与其他信号受体结合，仅响应配体 activation.这些观察结果表明， 细胞内的蛋白质在信号转导中起重要作用 转导，可能是负责AGE诱导的变化， 基因表达。