MEMBRANE LIPID PEROXIDATION AND SICKLE CELL PATHOLOGY

虽然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 在镰状细胞膜上发现了这种不可逆的 镰状细胞（ISC）。 这些ISC高度脱水， 变形性，改变的膜磷脂组织， 高凝状态和粘附于血管的倾向增加 内皮细胞 ISC的这些异常以及与 ISC计数和红细胞存活减少，结膜血管 异常和腿部溃疡的发生表明， 镰状细胞病发病机制中的ISC。 这个项目的目的是发现是否有任何相关性 膜脂过氧化程度、脂质不对称性改变 以及镰状细胞不同密度部分的高凝状态 在斯氏密度梯度上分离，这些膜是否 异常仅存在于ISC中，由于镰状化或 逐渐累积到膜损伤的异常。 体外细胞 由制霉菌素处理诱导的盘状镰状细胞的脱水将是 用于区分膜损伤与细胞损伤的主要作用， 高凝状态中的脱水本身，改变的膜脂质 镰状细胞的不对称性和过氧化膜脂质损伤。 本项目还将探讨外源性过氧化威胁是否在体外 与体内形成的ISC相比， 从镰状血中分离出来的 膜脂质过氧化分析的标准技术 将利用磷脂组织、凝固性和蛋白质， 在这个实验室里已经标准化了。 至少20个镰刀的数据 细胞患者将进行统计学分析。

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.