GLOMERULAR CAPILLARY WALL--NORMAL AND PATHOLOGIC

糖尿病肾病是糖尿病的主要并发症之一， 最终导致慢性肾衰竭。从形态学上讲， 其特征在于细胞外基质（ECM）的显著变化。 与这些变化相关的生化改变包括 IV型胶原蛋白表达增加， 蛋白聚糖（PG）。这些变化可以通过提高 实验模型系统中的己醛糖（葡萄糖）浓度，以及 认为与糖尿病患者蛋白尿有关 患者除了成年人的这些并发症， 青少年糖尿病患者的先天性异常的发生率增加 影响心脏、骨骼和神经系统以及泌尿生殖道， 也就是说，尾部退化综合征可以想象，这种进化 发育缺陷与高浓度的葡萄糖有关， 其扰乱形态发生调节因子的功能，即，ECM & 细胞粘附分子、生长因子、它们的受体和 原癌基因为了测试这种争论和可能的机制， 参与胚胎肾脏的器官发育异常，我们建议 通过使用一个 建立了小鼠后肾培养体系。 I.包括葡萄糖在内的己糖浓度升高对 胚胎肾的形态发生将通过以下方法进行监测： 形态测定分析、免疫组织化学和DNA复制研究。 随后，ECM蛋白质的从头合成和基因表达 （IV型胶原蛋白、层粘连蛋白和PG）和基质相关受体（整合素 α 3A、α 5、α 6B和β FGF-R）。生物化学， 免疫沉淀，翻译，转录，溶液 将使用杂交、RT-PCR和竞争性PCR方法。 二. 葡萄糖对细胞内ATP库磷酸肌醇的影响 代谢，以及参与酪氨酸磷酸化机制， 各种生长因子（IGF-I、胰岛素和EGF）的信号转导， 它们的受体（INS-R，IGF-IR）和原癌基因（c-fos，Egr，c-ret和c-fos） 罗斯）将进行调查。 三.最终，已知和未知的葡萄糖诱导/抑制 后肾中的转录物将被分离和表征 cDNA文库消减杂交和mRNA差异显示 方法.

In diabetes mellitus, nephropathy is one of the major complication that ultimately leads to chronic renal failure. Morphologically, it is characterized by remarkable changes in the extracellular matrices (ECM). The biochemical alterations which correlate with these changes include increased expression of type-IV collagen, & decreased synthesis of proteoglycans (PGs). These changes can be induced by elevating the aldohexose (glucose) concentrations in experimental model systems, and are believed to have some relationship with proteinuria in diabetic patients. in addition to these complications in adults, the offsprings of juvenile diabetics have increased incidence of congenital anomalies affecting heart, skeletal and nervous systems, and genitourinary tract, i.e., caudal regression syndrome. Conceivably, the evolution of such developmental defects is related to the high concentration of glucose, which perturbs the functions of morphogenetic regulators, i.e., ECM & cell adhesion molecules, growth factors, their receptors and protooncogenes. In order to test this contention & possible mechanisms involved in the dysorganogenesis of the embryonic kidneys, we propose to carry out experiments outlined under 3 major objectives by utilizing an established murine metanephric culture system. I. Effect of elevated concentrations of hexoses, including glucose, on the morphogenesis of the embryonic kidneys will be monitored by morphometric analyses, immunohistochemical and DNA replication studies. Following which, de novo synthesis and gene expressions of ECM proteins (type-IV collagen, laminin & PGs) & matrix related receptors (integrin alpha3A, alpha:5, alpha6B & betaFGF-R) will be investigated. Biochemical, immunoprecipitation, translational, transcriptional, solution hybridization, RT-PCR & competitive PCR methods will be used. II. Effect of glucose on the intracellular ATP stores, phosphoinositide metabolism, and the tyrosine phosphorylation mechanisms involved in the signal transduction of various growth factors (IGF-I, insulin & EGF), their receptors (INS-R, IGF-IR) & protooncogenes (c-fos, Egr, c-ret & c- ros) will be investigated. III. Eventually, the known and unknown glucose-induced/suppressed transcripts in the metanephric kidneys will be isolated and characterized by cDNA library subtractive hybridization and mRNA differential display methods.