TOXIC METABOLITES OF OXYGEN IN CATARACTOGENESIS

建议检验的假设是O2的活性物种是 氧化应激致晶状体血浆损伤的起始物 白内障的膜脂和膜蛋白。我们的目标是调查 白内障发病机制中涉及氧化的关键区域 蛋白质与质膜磷脂的过氧化。曝光 在体外对晶状体质膜进行各种氧化应激， 内源性膜蛋白MP26的修饰将被确定 采用SDS-聚丙烯酰胺凝胶电泳法。经过相同的治疗后， 膜磷脂的评估将通过测量形成 氨基磷脂、丙二醛加合物的薄层色谱、高效液相色谱和红外光谱研究 荧光分光光度法。脂肪酸将通过气相色谱和 高效液相色谱柱。丙二醛(MDA)和过氧化脂质将在 镜片观察它们与不饱和度损失的可能相关性 白内障中的脂肪酸。极谱和电子自旋共振波谱技术 将用于检测O2.-、OH、H_2O_2和不含阿霉素的半喹酮 房水中的自由基和添加小牛晶状体的兔晶状体中的自由基 微粒体NADPH氧化酶系统的体外研究。我们将尝试 聚合希夫碱偶联物的鉴定 晶状体中氨基磷脂在氧化应激和白内障中的作用 人和动物使用(14C-)丙二醛或磷脂酰乙醇胺。氧化 膜-SH在体外和前发性白内障中的应激交联性 而半乳糖诱导的兔早期白内障将通过以下方法确定 使用(14C-)碘乙酰胺。为了评估酶防御在细胞周期中的作用 白内障，超氧化物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。抗氧化剂的测试 在实验性白内障的治疗上还将继续。逮捕该名男子 人类早期白内障的进展将是巨大的 对病人的帮助。减税带来的经济效益 白内障手术的数量也不能被低估。

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.