TISSUE SPECIFIC NUTRITIONAL ADAPTATIONS IN RENAL FAILURE

肾衰竭时的组织特异性营养适应

• 批准号: 6635061
• 负责人: S. Russ Price
• 金额: $ 30.78万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6635061
• 关键词: acidosis; aldehyde /ketone oxidoreductase; aminoacid metabolism; branched chain aminoacid; cell line; chronic renal failure; diabetes mellitus; enzyme activity; gel mobility shift assay; genetic promoter element; glucocorticoids; high performance liquid chromatography; hormone regulation /control mechanism; insulin; kidney metabolism; laboratory rat; liver metabolism; muscle metabolism; northern blottings; nuclear runoff assay; nutrition related tag; phosphatidylinositol 3 kinase; protein kinase; transfection; western blottings

The essential branched-chain amino acids (BCAA) play critical roles in maintaining normal protein homeostasis and they influence critical intracellular signaling pathways that regulate metabolic functions. In normal individuals, nutritional adaptations to a reduced dietary protein intake (e.g., fasting, a low protein diet prescription) decrease the irreversible degradation of BCAA. Catabolic conditions like chronic renal failure (CRF) or acute diabetes impair these adaptive responses that preserve protein mass, thus contributing to the loss of lean body mass. The goals of Dr. Price and colleagues are to understand the mechanisms that regulate the activity of branched-chain alpha-ketoacid dehydrogenase (BCKAD), the rate-limiting enzyme in BCAA degradation, in the major tissues where BCAA are catabolized, and to determine if there are common signals in different catabolic states that regulate BCKAD activity, and hence, BCAA levels. To address these goals, the investigators will evaluate three hypotheses: 1) Acidification and glucocorticoids influence transcription of BCKAD subunit genes through specific cis-acting response elements. The investigators will identify specific DNA promoter elements in the BCKAD E2 gene that confer responses to acidification and glucocorticoids. 2) Abnormalities in BCAA utilization in rats with CRF result from tissue-specific alterations in BCKAD activity at both genetic and biochemical levels. The investigators will define how CRF influences the activities of BCKAD and BCKAD kinase, a unique kinase that inhibits BCKAD activity, in muscle, liver and kidney in a well-established rat model. They will measure BCKAD activity, BCKAD subunit and kinase proteins and amounts of subunit and kinase mRNAs 3) Insulin modulates BCKAD and/or BCKAD activities in different tissues by a mechanism requiring the critical signaling enzyme phosphatidylinositol 3-kinase. The investigators will determine the biochemical mechanism(s) that increase BCKAD activity in rat muscle, liver and kidney in response to acute diabetes mellitus (i.e., insulin insufficiency) and then examine the signaling mechanisms by which insulin regulates BCKAD and BCKAD kinase in cultured L6 muscle cells. The investigators findings will define cellular mechanisms regulating BCAA degradation in uremia, acute diabetes and other catabolic conditions.

必需支链氨基酸（BCAA）在人体内起着至关重要的作用。 在维持正常蛋白质稳态中的作用，它们影响着关键的 调节代谢功能的细胞内信号通路。正常 个体，对减少的膳食蛋白质摄入的营养适应（例如， 禁食，低蛋白饮食处方）减少不可逆降解 的BCAA。分解代谢疾病，如慢性肾衰竭（CRF）或急性 糖尿病损害了这些保持蛋白质质量的适应性反应， 导致瘦体重的减少。普赖斯博士的目标 同事们要了解的机制，调节活动的 支链α-酮酸脱氢酶（BCKAD），限速酶 在BCAA降解中，在BCAA被分解代谢的主要组织中， 确定在不同的分解代谢状态下是否有共同的信号， 调节BCKAD活性，从而调节BCAA水平。为了实现这些目标， 研究人员将评估三个假设：1）酸化和 糖皮质激素影响BCKAD亚基基因转录的特异性途径 顺式作用反应元件。调查人员将确定特定的DNA BCKAD E2基因中赋予酸化反应的启动子元件 和糖皮质激素。2)慢性肾衰竭大鼠支链氨基酸利用的异常 结果从组织特异性改变BCKAD活性在遗传和 生化水平。研究者将定义CRF如何影响 BCKAD和BCKAD激酶（一种抑制BCKAD的独特激酶）的活性 活性，在肌肉，肝脏和肾脏在一个完善的大鼠模型。他们 将测量BCKAD活性、BCKAD亚基和激酶蛋白以及 3）胰岛素调节BCKAD和/或BCKAD活性， 不同的组织通过一种需要关键信号酶的机制 磷脂酰肌醇3-激酶调查人员将确定 增加大鼠肌肉、肝脏和肾脏中BCKAD活性的机制 对急性糖尿病的反应（即，胰岛素不足），然后 研究胰岛素调节BCKAD和BCKAD的信号传导机制 在培养的L 6肌细胞中的激酶。调查结果将确定 尿毒症、急性糖尿病和糖尿病患者中调节支链氨基酸降解的细胞机制 其他代谢条件。