STUDIES OF HEMOLYTIC ANEMIA

溶血性贫血的研究

• 批准号: 3341066
• 负责人: DAVID J. JOLLOW
• 金额: $ 15.28万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-07-01 至 1993-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3341066
• 关键词: aniline; ankyrins; blood toxicology; cell senescence; covalent bond; dapsone; dosage; drug adverse effect; drug design /synthesis /production; drug metabolism; erythrocytes; free radical oxygen; glucose 6 phosphate dehydrogenase deficiency; hemolysis; hemolytic anemia; hemotoxin; high performance liquid chromatography; laboratory rat; membrane lipids; membrane proteins; methemoglobin; oxidation; primaquine; radiation detector; radiotracer; scintillation spectrometry; spectrin; spectrometry; stainings; stoichiometry; thin layer chromatography; toxicant interaction; toxin metabolism

A profound hemolytic anemia following exposure to arylamine drugs and environmental chemicals affects Americans of African and Mediterranean descent (Italian, Greek, etc.). In the 50's and 60's, extensive studies on primaquine and phenylhydrazine led to the concept that (re) active metabolites of the drugs damaged red cells by "oxidative stress". The hemotoxic metabolites however were not identified and the postulate of "oxidative stress" as the mechanism underlying drug-induced hemolytic anemia was not further developed. We have now identified hemolytic metabolites of aniline, dapsone and phenacetin and observed high hemolytic activity with a putative metabolite of primaquine. Mechanism studies using the hemolytic metabolite of depsone (DDS-NOH) in rat red cells indicated that DDS-NOH causes a), formation of disulfide-linked adducts between certain membrane skeletal proteins and hemoglobin monomers, and b), development of an extreme echinocyte morphology. These data, taken together with recent literature observations on acitive oxygen biochemistry, red cell skeletal protein function, and the normal mechanisms of splenic sequestration of "old" red cells, suggest an overall working hypothesis for the mechanism of drug-induced hemolytic anemia; viz that the toxic drug metabolites interact with oxyhemoglobin to generate oxygen free radicals, which in turn generate free radicals of hemoglobin sulfhydryl groups. Adduct formation between hemoglobin and membrane skeletal proteins results in loss of stuctural integrity and activation of senescent antibody binding sites on the cell surface, with subsequent macrophage phagocytosis. The objectives of the present studies are a), to indentify the hemolytic metobolites of primaquine, b) to define the role of acitve oxygen species in activating red cell protein sulfhydryl groups and in the commitment of DDS-NOH damaged red cells to splenic sequestration, c) to identify the domains of spectrin andd ankyrin to which the hemoglobin monomers bind and to assess the structural significance of this binding, and d), to determine whether or not the senescent antibody plays a role in the removal of DDS-NOH damaged red cells from the circulation. It is expected that these studies will determine the relative importance of N vs C oxidation products of primaquine in primaquine hemotoxicity, establish whether or not active oxygen species are obligatory in the hemolytic sequence, and provide a firm basis for more detailed resolution of how skeletal protein structure on the inside of the red cell membrane determines the properties and structures of the outer surface of the red cell.

暴露于芳胺后发生严重溶血性贫血 毒品和环境化学物质影响着非洲的美国人， 和地中海血统（意大利，希腊等）。 在50年代和 60年代，伯氨喹和苯肼的广泛研究导致 药物的活性代谢物破坏红细胞的概念 细胞的“氧化应激”。 然而， 没有被确定，“氧化应激”的假设是 药物诱导的溶血性贫血的潜在机制不是 进一步发展。 我们已经鉴定出溶血代谢物 苯胺，氨苯砜和非那西丁，并观察到高溶血性 与伯氨喹的推定代谢物的活性。 机制 使用Depsone的溶血代谢物（DDS-NOH）的研究 大鼠红细胞表明，DDS-NOH导致a），形成 某些膜骨架之间的二硫键连接的加合物 蛋白质和血红蛋白单体，和B）， 极端的棘细胞形态。 这些数据与 活性氧生物化学的最新文献观察 细胞骨架蛋白功能，以及 脾隔离的“老”红细胞，表明一个整体的工作 药物性溶血性贫血机制假说; 即毒性药物代谢物与氧合血红蛋白相互作用 产生氧自由基，这反过来又产生自由基， 血红蛋白巯基的自由基。 加合物形成 血红蛋白和膜骨架蛋白之间的联系 结构完整性丧失和衰老抗体活化 细胞表面的结合位点，随后巨噬细胞 吞噬作用 本研究的目标是：（a） 鉴定伯氨喹的溶血代谢物，B）确定 活性氧在活化红细胞蛋白中作用 巯基和在DDS-NOH的承诺受损 红细胞脾隔离，c）以确定 血影蛋白和锚蛋白，血红蛋白单体与之结合， 评估这种结合的结构意义，和d）， 确定衰老抗体是否在 DDS-NOH从循环中去除受损的红细胞。 预计这些研究将确定相对 伯氨喹的N与C氧化产物在 伯氨喹血液毒性，确定是否有活性氧 种在溶血序列中是必须的，并提供了一个 为更详细地解决骨骼蛋白质 红细胞膜内部的结构决定了 红细胞外表面的性质和结构。