TOXIC METABOLITES OF OXYGEN IN CATARACTOGENESIS

白内障发生中氧的有毒代谢物

• 批准号: 3257331
• 负责人: KAILASH C BHUYAN
• 金额: $ 22.39万
• 依托单位: MOUNT SINAI SCHOOL OF MEDICINE OF CUNY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-09-01 至 1993-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3257331
• 关键词: Schiff bases; aldehyde reductase; antioxidants; ascorbate; catalase; cataract; chemical conjugate; deferoxamine; drug screening /evaluation; electron spin resonance spectroscopy; eye agent; eye disorder chemotherapy; fluorescence spectrometry; free radicals; gel electrophoresis; glutathione peroxidase; high performance liquid chromatography; human therapy evaluation; human tissue; hydrogen peroxide; hydroxyl group; laboratory mouse; laboratory rabbit; laboratory rat; lens proteins; lipid peroxides; membrane lipids; membrane proteins; molecular pathology; oxidation; peroxidation; phospholipids; polarography; polymerization; radioimmunoassay; respiratory oxygen; superoxide dismutase; thin layer chromatography; unsaturated fatty acids

The hypothesis proposed to test is that reactive species of O2 are the initiators of oxidative stress-induced damage to the lenticular plasma membrane lipids and proteins in cataract. The objective is to investigate the crucial areas of the mechanism of cataract involving oxidation of proteins and peroxidation of phospholipids of plasma membranes. Exposing the lens plasma membranes to various oxidative stresses in vitro, modification of the intrinsic membrane protein MP26, will be determined using SDS polyacrylamide gel electrophoresis. After identical treatment, membrane phospholipids will be assessed by measuring the formation of aminophospholipid.malondialdehyde adduct using TLC, HPLC and spectrofluorometry. Fatty acids will be analysed by gas chromatography and HPLC. Malondialdehyde(MDA) and lipid hydroperoxides will be estimated in lens to observe their possible correlation with the loss of unsaturated fatty acids in cataracts. Polarographic and ESR spectroscopic techniques will be used to detect O2.-, OH., H2O2 and adriamycin semiquinone free radical in aqueous humor, and in lens of rabbit by addition of calf lens microsomal NADPH oxidase system in vitro. Attempts will be made to identify Schiff base conjugate formed by polymerization of aminophospholipids of lens under oxidative stress and in cataracts in the human and animal using (14C-)MDA or phosphatidyl ethanolamine. Oxidative stress-induced cross linking of membrane-SH in vitro, and in precataract and early cataract induced in rabbits by galactose will be determined by using (14C-)iodoacetamide. To assess the role of enzymic defenses in cataracts, analysis of the bioactive and inactive forms of superoxide dismutase, catalase and GSH peroxidase will be done using spectrophotometric technique and radioimmunoassay. Tests of antioxidants in the therapy of experimental cataracts will be continued. Arrest of the progression of cataract in the human at an early stage will be of immense help to the patients. The economic benefit resulting from reduction of the number of surgical procedures for cataracts also must not be underestimated.

提出要检验的假设是 O2 的活性物质是 氧化应激引起的透镜状等离子体损伤的引发剂 白内障中的膜脂和蛋白质。 目的是调查 白内障机制的关键领域涉及氧化 蛋白质和质膜磷脂的过氧化。 暴露 晶状体质膜在体外对各种氧化应激的影响， 内在膜蛋白 MP26 的修饰，将被确定 采用SDS聚丙烯酰胺凝胶电泳。 经过同样的处理后， 将通过测量膜磷脂的形成来评估 氨基磷脂.丙二醛加合物，使用 TLC、HPLC 和 荧光分光光度法。 脂肪酸将通过气相色谱法进行分析 高效液相色谱法。 丙二醛（MDA）和脂质氢过氧化物将在 透镜观察它们与不饱和度损失可能的相关性 白内障中的脂肪酸。 极谱和 ESR 光谱技术 将用于检测不含 O2.-、OH.、H2O2 和阿霉素半醌 房水中的自由基，以及通过添加小牛晶状体在兔子的晶状体中的自由基 体外微粒体 NADPH 氧化酶系统。 将努力 鉴定由聚合形成的希夫碱缀合物 氧化应激下和白内障中晶状体的氨基磷脂 人类和动物使用 (14C-)MDA 或磷脂酰乙醇胺。 氧化性 体外和白内障前期应激诱导的膜-SH 交联 半乳糖引起的兔子早期白内障将通过以下方法确定 使用(14C-)碘乙酰胺。 评估酶防御的作用 白内障，超氧化物的生物活性和非活性形式的分析 歧化酶、过氧化氢酶和谷胱甘肽过氧化物酶将使用 分光光度技术和放射免疫分析。 抗氧化剂测试 实验性白内障的治疗将继续进行。 逮捕 人类白内障在早期阶段的进展将是巨大的 帮助患者。 减少所产生的经济效益 白内障手术的数量也不容低估。