ERYTHROCYTE SENESCENCE

红细胞衰老的机制仍是一个悬而未决的问题。连 尽管对基因决定的溶血性贫血的研究 显示出各种缺陷会缩短红细胞寿命 斯潘，关于正常的机制，还没有出现明确的答案 测定红细胞存活率。红细胞衰老的检测 由于分离老化细胞的困难而受到阻碍。各种各样的 已经利用技术来获得这些衰老的红细胞 但大多数情况并不令人满意。该提案描述了一种新的程序，用于 兔衰老红细胞的分离。有了这个 方法：用生物素共价标记兔红细胞 与N-羟基琥珀酰亚胺生物素反应。结合的生物素已被证明 在输注衍生细胞时不影响红细胞存活 进入兔子体内。在输液后的不同时间，生物素 亲和素亲和力可选择性回收红细胞 层析法。在这个过程中，红细胞通常被分离出来。 在预期死亡后的5至10天内。这些衰老的红细胞 已发现其5‘-三磷酸腺苷水平正常。这个 目前的提案将研究这些衰老的其他特征 红细胞包括糖酵解的完整性和 己糖单磷酸途径，与腺嘌呤核苷酸相关的酶 新陈代谢，细胞抗氧化防御的状态， 细胞表面的抗体和骨骼/膜的完整性 很复杂。仔细检查这些参数应该会带来更好的 对控制黄瓜正常衰老因素的认识 红血球。

The mechanism of erythrocyte senescence remains an unsolved problem. Even though the study of genetically determined hemolytic anemias has demonstrated a variety of defects which can shorten the erythrocyte life span, no clear cut answer has emerged as to the normal mechanisms which determine erythrocyte survival. The examination of erythrocyte senescence has been hindered by the difficulty of isolating aged cells. A variety of techniques have been utilized for obtaining these senescent erythrocytes but most are not satisfactory. This proposal describes a new procedure for the isolation of senescent erythrocytes from the rabbit. With this technique, rabbit erythrocytes are covalently labeled with biotin by reaction with N-hydroxysuccinimido biotin. The bound biotin has been shown to not affect erythrocyte survival when the derivatized cells are infused into the rabbit. At various times after infusion, the biotinylated erythrocytes can be selectively recovered by avidin affinity chromatography. With this procedure, erythrocytes are routinely isolated within 5 to 10 days of their expected death. These senescent erythrocytes have been found to have normal levels of adenosine 5' - triphosphate. The present proposal will examine additional characteristics of these senescent erythrocytes including the integrity of the glycolytic and hexosemonophosphate pathways, the enzymes related to adenine nucleotide metabolism, the state of the cellular anti-oxidant defenses, the level of antibodies on the cell surface and the integrity of the skeleton/membrane complex. A careful examination of these parameters should lead to a better understanding of factors which control the normal senescence of erythrocytes.