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红细胞衰老会导致大鼠红细胞表面特性发生改变,但人类红细胞不会。通过在双聚合物水相系统中分配进行研究。

Aging of erythrocytes results in altered red cell surface properties in the rat, but not in the human. Studies by partitioning in two-polymer aqueous phase systems.

作者信息

Walter H, Krob E J, Ascher G S

出版信息

Biochim Biophys Acta. 1981 Feb 20;641(1):202-15. doi: 10.1016/0005-2736(81)90584-8.

Abstract

Aqueous solutions of dextran and of poly(ethylene glycol) when mixed give rise to two-phase systems useful in separating cells, on the basis of their surface properties, by partitioning. Depending on whether salts with unequal or equal affinity for the two phases are chosen, phases with or without an electrostatic potential difference between the phases are obtained. At appropriate polymer concentrations the former yield cell partition coefficients (i.e., the quantity of cells in the top phase as a percentage of total cells added) based on charge-associated surface properties while the latter reflect membrane lipid-related parameters. With increasing cell age, rat erythrocytes have diminishing partition coefficients in both charged and uncharged phases. Using the elevated aspartate aminotransferase levels of younger red cells as a marker, we have not found that young mature erythrocytes of human do not have the highest partition coefficient in the red cell population as they do in rat. Experiments with isotopically labeled dog red cells yield results similar to those found with human erythrocytes. Furthermore, density-separated young and old red cells from human give overlapping countercurrent distribution curves. Finally, countercurrent distribution of human red blood cells followed by pooling of cells from the left and right ends of the distribution and subjection of these cells to a redistribution gives curves that overlap with each other and with the original countercurrent distribution. This indicates that not only are human red cells not subfractionated based on possible age-related surface alterations, but also that they are not subfractionated by partitioning based on any surface parameter. These results are consistent with our previous findings that membrane sialic acid/hemoglobin absorbance is essentially constant through the extraction train after countercurrent distribution of human erythrocytes in a charged phase system; and with the recent reports of others that there is no difference in electrophoretic mobility between human young and old red cells.

摘要

葡聚糖和聚乙二醇的水溶液混合时会形成两相系统,该系统可根据细胞的表面性质通过分配作用来分离细胞。根据选择的盐对两相的亲和力是否不等,可得到两相之间有或没有静电势差的相。在合适的聚合物浓度下,前者基于与电荷相关的表面性质产生细胞分配系数(即上层相中细胞数量占添加的总细胞数量的百分比),而后者反映与膜脂质相关的参数。随着细胞老化,大鼠红细胞在带电和不带电相中分配系数都逐渐降低。以年轻红细胞中升高的天冬氨酸转氨酶水平作为标志物,我们并未发现人类年轻成熟红细胞在红细胞群体中不像大鼠那样具有最高的分配系数。用同位素标记的犬红细胞进行的实验得到的结果与人类红细胞相似。此外,对人类密度分离的年轻和年老红细胞进行逆流分布实验,得到的曲线相互重叠。最后,对人类红细胞进行逆流分布,然后将分布两端的细胞合并并重新进行分配,得到的曲线相互重叠且与原始逆流分布曲线重叠。这表明人类红细胞不仅不会基于可能与年龄相关的表面改变进行亚分级,而且也不会基于任何表面参数通过分配作用进行亚分级。这些结果与我们之前的发现一致,即在带电相系统中对人类红细胞进行逆流分布后,整个提取过程中膜唾液酸/血红蛋白吸光度基本恒定;也与其他人最近的报道一致,即人类年轻和年老红细胞的电泳迁移率没有差异。

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