MEMBRANE LIPID PEROXIDATION AND SICKLE CELL PATHOLOGY

尽管镰状细胞病是一种与异常血红蛋白有关的疾病 在红细胞内，各种形态和功能异常 在镰状细胞的膜中被鉴定为不可逆转的 镰状细胞(ISC)。这些ISC高度脱水，并已减少 易变形，膜磷脂结构改变， 高凝状态，与血管粘连的倾向增加 内皮细胞。ISC的这些异常以及两者之间的关系 ISC计数与红细胞存活率、结膜血管 异常和腿部溃疡的发生表明 ISC在镰状细胞病发病机制中的作用 这个项目的目的是发现两者之间是否存在任何关联 膜脂过氧化程度、脂质不对称性改变 和不同密度组分的镰状细胞的高凝状态 分离出的斯特拉坦密度梯度和这些膜是否 异常只出现在ISC中，原因是镰刀或 逐渐增加到膜损伤的异常。体外细胞 制霉菌素治疗引起的盘状镰状细胞脱水 用于区分膜损伤和细胞损伤的主要作用 脱水本身在高凝状态下，改变了膜脂 镰状细胞的不对称性和过氧化损伤。 该项目还将探索外源性过氧化物质在体外是否会威胁到 盘状镰状细胞可产生与体内形成的ISC相当的ISC 从镰刀血中分离出来的。 膜脂过氧化分析的标准技术 将利用磷脂组织、凝固性和蛋白质 已经在这个实验室里标准化了。来自至少20把镰刀的数据 对细胞病人进行统计分析。

Although sickle cell disease is a disease related to abnormal hemoglobin iside the red cell, a variety of morphologcal and functional abnormalities have been identified in the membranes of sickle cells called irreversibly sickled cells (ISC). These ISC are highly dehydrated and have reduced deformability, altered membrane phospholipid organization, hypercoagulability, and increased tendency to adhere to vascular endothelium. These abnormalities of ISC as well as a relationship between ISC counts and reduced red cell survival, conjunctival blood vessel abnormalities, and occurrence of leg ulcers suggest a potential role for ISC in the pathogenesis of sickle cell disease. The aim of this project is to discover whether there is any correlation between the degree of membrane lipid peroxidaition, altered lipid asymmetry and hypercoagulability in varying density fractions of sickle cells separated on Stractan density gradients and whether these membrane abnormalities are present only in the ISC due to sickling or are abnormalities that accrue gradually to membrane damage. In vitro cellular dehydration induced by Nystatin treatment to discoid sickle cells will be used to discriminate predominant role of membrane damage versus cellular dehydration per se in the hypercoagulability, altered membrane lipid asymmetry and peroxidative membrane lipid damage of sickle cells. This project also will explore whether exogenous peroxidant threat in vitro to discoid sickle cells can generate ISC comparable to in vivo-formed ISC fractionated from sickle blood. Standard techniques for analyses of membrane lipid peroxidation, membrane phospholipid organization, coagulability, and proteins will be utilized and have been standardized in this laboratory. Data from at least 20 sickle cell patients will be analyzed statistically.