METABOLIC STRESS IN NORMAL AND ABNORMAL HUMAN RED CELLS

正常和异常人类红细胞的代谢应激

• 批准号: 3136743
• 负责人: EUGENE F ROTH
• 金额: $ 9.03万
• 依托单位: MONTEFIORE MEDICAL CENTER (BRONX, NY)
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-01-01 至 1988-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3136743
• 关键词: amidophosphoribosyltransferase; antioxidants; blood /lymphatic pharmacology; density gradient ultracentrifugation; enzyme mechanism; erythrocyte membrane; erythrocytes; glucose 6 phosphate dehydrogenase deficiency; glutathione; glycolysis; host organism interaction; human tissue; lipid peroxides; liposomes; lysine; malaria; membrane lipids; microorganism growth; microorganism metabolism; myelinopathy; oxidation; pentose phosphate shunt; radiotracer; stress; tissue /cell culture; virulence

In human red cells, stress occurs when the normally slow moving metabolic machinery of the red cell is forced to accelerate beyond its normal rate. Metabolic stress is found in red cells parasitized by malaria and in some pathological states such as in myeloproliferative diseases (MPD). Glucose-6-phosphate dehydrogenase deficient red cells (G6PD-) are especially vulnerable to stress because they cannot augment pentose shunt metabolism. This study intends to investigate the salient features of normal and abnormal human red cells infected in vitro with falciparum malaria. In particular, the mechanism whereby G6PD- cells inhibit the growth of malaria will be studied. Two hypotheses will be tested: A. Oxidative stress in malaria infected cells interacts with the enzyme deficiency to inhibit growth. B. G6PD- cells cannot produce enough ribose phosphate for nucleotide needs of the growing parasites. These two hypotheses are not mutually exclusive. Methodology will include culture of malaria in vitro, assays for susceptibility to membrane lipid peroxidation, incorporation of labelled precursors into nucleotides, and a study of the growth of malaria in relation to the supply of phosphoribosylpyrophosphate (PRPP) - a key metabolite of the pentose pathway. MPD red cells are characterized by high concentrations of reduced glutathione (GSH) and increased glycolytic enzyme activity and in some ways resemble immature red cells or cord cells. They also have an increased susceptibility to membrane lipid peroxidation, which is linked somehow to the high GSH content. The nature of this relationship will be explored by studying the tendency of crude and purified red cell lysates to oxidize hemoglobin and elaborate active forms of 02. The effect of these lysates on sealed red cell ghosts and liposomes will also be studied. GSH may function as a pro-oxidant as well as an anti-oxidant under some conditions. Whether or not pro-oxidant conditions occur in MPD red cells remains to be determined. The requirement of PRPP synthetase for GSH in intact red cells will also be studied as will GSH synthesis and degradation rates. Manipulations of GHS may alter the course of MPD, reduce transfusion needs, inprove blood storage, enhance knowledge.

在人类红细胞中，当正常缓慢移动的代谢产物 红细胞的机械被迫加速超过其正常速率。 代谢应激存在于被疟疾寄生的红细胞中， 病理状态，如骨髓增生性疾病（MPD）。 葡萄糖-6-磷酸脱氢酶缺陷型红细胞（G6 PD-） 特别容易受到压力，因为它们不能增加戊糖分流 新陈代谢. 本研究旨在探讨 体外感染恶性疟原虫的正常和异常人红细胞 疟疾 特别是，G6 PD-细胞抑制肿瘤细胞增殖的机制， 将研究疟疾的增长情况。 两个假设将被测试：A。 疟疾感染细胞中的氧化应激与酶相互作用 缺乏抑制生长。 B。G6 PD-细胞不能产生足够的核糖 磷酸盐来满足寄生虫生长所需的核苷酸。 这两 假设并不相互排斥。 方法将包括文化 体外疟疾，膜脂质过氧化敏感性测定， 将标记的前体掺入核苷酸，以及对 疟疾的增长与磷酸核糖焦磷酸的供应有关 （PRPP）-戊糖途径的关键代谢物。 MPD红细胞是 特征在于高浓度的还原型谷胱甘肽（GSH）， 糖酵解酶活性增加，在某些方面类似于未成熟红 细胞或脐带细胞。 他们也有一个增加的易感性， 膜脂质过氧化，这与高GSH有某种联系 内容 这种关系的性质将通过研究 粗品和纯化的红细胞裂解物氧化血红蛋白的趋势， 02的详细活动形式。 这些裂解物对密封红细胞的影响 还将研究细胞血影和脂质体。 GSH可以作为一种 在某些条件下是促氧化剂和抗氧化剂。 是否 MPD红细胞中不发生促氧化条件仍有待进一步研究。 测定 完整红细胞中PRPP合成酶对GSH的需求 还将研究GSH的合成和降解速率。 GHS的操作可以改变MPD的过程，减少输血需求， 改善血液储存，增加知识。