BILIRUBIN ENCEPHALOPATHY--MODELS AND METABOLIC STUDIES

胆红素脑病——模型和代谢研究

• 批准号: 3081432
• 负责人: WILLIAM RHINE
• 金额: $ 7.95万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3081432
• 关键词: blood brain barrier; disease /disorder model; hemolysis; hyperbilirubinemia; kernicterus; laboratory rabbit; magnetic resonance imaging; membrane permeability; metalloporphyrins; model design /development; nervous system disorder; neuroblastoma; nuclear magnetic resonance spectroscopy; tissue /cell culture

The prime objective of this proposal is to development of the candidate's skills in basic research while gaining insight into the pathophysiology and mechanisms of kernicterus and bilirubin toxicity. By coursework, discussions and supervised investigation, understanding will be gained in cellular metabolism and physiology at a level particularly appropriate for a clinical investigator in neonatology and developmental biology. The scientific objective of this proposal is aimed at understanding the still obscure underlying cellular injuries fundamental kernicterus. Kernicterus, although primarily associated with erythroblastosis fetalis previously, has been found in 15-25% of autopsied newborns. The development of kernicterus depends on not only on serum level of unconjugated bilirubin; apparently binding to albumin and integrity of the blood-brain barrier are at least two mitigating factors affecting bilirubin injury. Prior models have been developed to determine what are the characteristic histopathologic and behavioral changes of kernicterus. However, previous studies including our own have shown that some of these models produce only a transient bilirubin encephalopathy (TBE) in these animals. In vitro, bilirubin has been shown to be toxic to many cellular functions of many different types of cells. Which of these toxicities predominates in kernicterus is known; determining this depends upon demonstration of comparable physiologic dysfunction in both in vivo and in vitro models. This proposed research plan initially develops and characterizes different animal models of kernicterus, including a hemolysis-induced model simulating hemolytic conditions associated with kernicterus. Histopathology and behavioral observations will define kernicterus injury. Kernicterus injury will then be characterized by neurophysiology, magnetic resonance imaging and spectroscopy studies, including evaluation of energy metabolism changes. Using these physiologic and metabolic markers, similarities and distinction between kernicterus and TBE will be studied. In vivo studies will also examine therapies to decrease bilirubin production and their ability to prevent kernicterus. In vitro studies will further evaluate bilirubin toxicity on neural-derived cells. Magnetic resonance spectroscopy studies of energy metabolism will allow direct comparison to in vivo animal models of kernicterus and the brain energy metabolism changes seen by our planned studies. This will help validate the use of this model for further studies of bilirubin toxicity. Subsequent studies will evaluate cellular environment including pH and albumin concentration, as it affects bilirubin toxicity in these cells. Finally, investigation of bilirubin toxicity will include assaying effects on membrane physiology of ion channels. These channels are well- characterize in neuroblastoma cells but not as affected by bilirubin. If necessary, contingencies are planned to develop alternate cell lines for in vitro studies of bilirubin toxicity.

该提案的主要目标是发展候选人的能力 基础研究技能，同时深入了解病理生理学和 核黄疸和胆红素毒性的机制。 通过课程作业， 讨论和监督调查，将获得理解 细胞代谢和生理学水平特别适合 新生儿学和发育生物学的临床研究者。 该提案的科学目标旨在了解 仍然掩盖了根本性核黄疸的潜在细胞损伤。 核黄疸，虽然主要与胎儿成红细胞增多症有关 此前，已在 15-25% 的新生儿尸检中发现了这种情况。 这 核黄疸的发生不仅取决于血清中 未结合胆红素；明显与白蛋白结合并保持其完整性 血脑屏障是影响胆红素的至少两个缓解因素 受伤。 先前的模型已经被开发来确定什么是 核黄疸的特征性组织病理学和行为变化。 然而，包括我们自己在内的先前研究表明，其中一些 模型仅产生短暂性胆红素脑病（TBE） 动物。 在体外，胆红素已被证明对许多细胞具有毒性。 许多不同类型细胞的功能。 这些毒性中的哪一个 已知在核黄疸中占主导地位；确定这一点取决于 证明体内和体内具有可比较的生理功能障碍 体外模型。 这项拟议的研究计划最初开发并描述了不同的 核黄疸动物模型，包括溶血诱导模型 模拟与核黄疸相关的溶血情况。 组织病理学和行为观察将确定核黄疸损伤。 核黄疸损伤将通过神经生理学、磁性 共振成像和光谱研究，包括能量评估 新陈代谢的变化。 利用这些生理和代谢标志物， 将研究核黄疸和 TBE 之间的异同。 体内研究还将检查降低胆红素的疗法 生产及其预防核黄疸的能力。 体外研究将进一步评估胆红素对神经源性细胞的毒性 细胞。 能量代谢的磁共振波谱研究将 允许与核黄疸的体内动物模型进行直接比较 我们计划的研究发现大脑能量代谢发生变化。 这将 帮助验证该模型在胆红素进一步研究中的用途 毒性。 后续研究将评估细胞环境，包括 pH 值和白蛋白浓度，因为它会影响这些物质中的胆红素毒性 细胞。 最后，胆红素毒性的调查将包括测定 对离子通道膜生理学的影响。 这些渠道都很好—— 神经母细胞瘤细胞的特征，但不受胆红素的影响。 如果 必要时，计划开发替代细胞系 胆红素毒性的体外研究。

项目成果

Gd-DTPA MR detection of blood-brain barrier opening in rats after hyperosmotic shock.

Gd-DTPA MR检测大鼠高渗休克后血脑屏障的开放情况。

• DOI: 10.1097/00004728-199307000-00008
• 发表时间: 1993
• 期刊: Journal of computer assisted tomography
• 影响因子: 1.3
• 作者: Rhine,WD;Benaron,DA;Enzmann,DR;Chung,C;Gonzales-Mendez,R;Sayre,JR;Stevenson,DK
• 通讯作者: Stevenson,DK

Bilirubin toxicity and differentiation of cultured astrocytes.

• DOI: 10.1038/sj.jp.7200180
• 发表时间: 1999-04-01
• 期刊: Journal of perinatology : official journal of the California Perinatal Association
• 作者: Rhine, W D;Schmitter, S P;Stevenson, D K
• 通讯作者: Stevenson, D K