GLOMERULAR CAPILLARY WALL--NORMAL AND PATHOLOGIC

肾小球毛细血管壁——正常和病理

• 批准号: 2138172
• 负责人: Yashpal S. Kanwar
• 金额: $ 26.2万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-04-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2138172
• 关键词: DNA replication; biological signal transduction; collagen; diabetic nephropathy; extracellular matrix; gene expression; genetic translation; glomerular filtration; growth factor; hexoses; immunocytochemistry; in situ hybridization; integrins; laboratory mouse; laboratory rabbit; laboratory rat; mammalian embryology; medical complication; membrane structure; molecular pathology; phosphatidylinositols; polymerase chain reaction; tissue /cell culture

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.

在糖尿病中，肾病是糖尿病的主要并发症之一 最终导致慢性肾功能衰竭。从形态上讲，它是 以细胞外基质(ECM)的显著变化为特征。 与这些变化相关的生化变化包括 IV型胶原表达增加，合成减少 蛋白多糖(PG)。这些变化可以通过提升 实验模型体系中的乙醛已糖(葡萄糖)浓度，以及 被认为与糖尿病患者的蛋白尿有关 病人。除了成人的这些并发症外，后代 的青少年糖尿病患者增加了先天性畸形的发生率 影响心脏、骨骼和神经系统，以及泌尿生殖道， 即尾部退缩综合征。可以想象，这样的进化 发育缺陷与葡萄糖浓度过高有关， 这扰乱了形态发生调节因子的功能，即ECM& 细胞黏附分子、生长因子及其受体和 原癌基因。为了测试这一争论和可能的机制 参与胚胎肾脏的器官发育障碍，我们建议 通过利用一个 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 胚胎肾脏的形态发生将通过以下方式进行监测 形态计量分析、免疫组织化学和DNA复制研究。 随后，细胞外基质蛋白的从头合成和基因表达 (IV型胶原、层粘连蛋白和前列腺素)与基质相关受体(整合素 阿尔法3A、阿尔法：5、阿尔法6B和贝塔成纤维细胞生长因子-R)。生化的， 免疫沉淀，翻译，转录，溶液 将采用杂交、RT-PCR法和竞争性PCR法。 II.葡萄糖对细胞内ATP储备--肌醇磷脂的影响 代谢，以及参与的酪氨酸磷酸化机制 多种生长因子(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- ROS)将接受调查。 最终，已知和未知的葡萄糖诱导/抑制 后肾中的转录本将被分离和鉴定 用文库消减杂交和mRNA差异显示 方法：研究方法。