Flatman P W, Smith L M
Department of Physiology, University Medical School, Teviot Place, Edinburgh EH8 9AG, UK.
Pflugers Arch. 1996 Oct;432(6):995-1002. doi: 10.1007/s004240050227.
Mg efflux from ferret red blood cells is stimulated when cells are Mg loaded, but the properties of efflux depend on the loading method. When cell Mg content is altered using A23187, which is subsequently washed away, Mg efflux is minimal until intracellular ionized [Mg] ([Mg2+]i) is greater than 0.9 mM, it then increases substantially with [Mg2+]i. Efflux from loaded cells falls as external [Na] ([Na]o) is reduced, and net Mg influx (against an electrochemical gradient) is seen when [Na]o is sufficiently low. Both influx and efflux are amiloride sensitive. Mg influx from media containing a normal or low [Na] is not affected by reducing [Mg2+]i to very low levels. When cells are Mg loaded by incubating them in media containing 5 mM Na and Mg, Mg efflux is again minimal until [Mg2+]i is greater than 0.9 mM and then it increases with [Mg2+]i, but at a rate approximately 4 times faster than in cells loaded using A23187. This efflux is little affected by 1 mM amiloride. Thus Mg-loading using A23187 reveals the [Mg2+]i dependence of a transporter which is amiloride sensitive, reversible and can operate against an electrochemical gradient, consistent with Na-Mg antiport. Loading by incubation in low-[Na] media activates a high-capacity Mg transporter which obscures the antiporter.
当雪貂红细胞镁负荷增加时,镁外流受到刺激,但外流特性取决于加载方法。当使用随后被洗脱的A23187改变细胞镁含量时,在细胞内游离镁浓度([Mg2+]i)大于0.9 mM之前,镁外流极少,之后随着[Mg2+]i升高而显著增加。加载细胞的外流随着细胞外钠浓度([Na]o)降低而减少,当[Na]o足够低时会出现净镁内流(逆电化学梯度)。内流和外流均对氨氯地平敏感。从含有正常或低[Na]的培养基中镁内流不受将[Mg2+]i降至极低水平的影响。当细胞在含有5 mM Na和Mg的培养基中孵育进行镁加载时,在[Mg2+]i大于0.9 mM之前,镁外流同样极少,之后随[Mg2+]i增加,但速率比使用A23187加载的细胞快约4倍。这种外流几乎不受1 mM氨氯地平的影响。因此,使用A23187进行镁加载揭示了一种对氨氯地平敏感、可逆且能逆电化学梯度运行的转运体对[Mg2+]i的依赖性,这与钠-镁反向转运一致。在低[Na]培养基中孵育加载激活了一种高容量镁转运体,从而掩盖了反向转运体。