Locher R, Neyses L, Stimpel M, Küffer B, Vetter W
Biochem Biophys Res Commun. 1984 Nov 14;124(3):822-8. doi: 10.1016/0006-291x(84)91031-3.
In order to study the influence of the cholesterol content on the calcium entry channel, the human red blood cell was used as a model system. The cholesterol to lecithin ratio (C/L ratio) of the membrane was modified experimentally by incubating the cells (15h, 25 degrees) with liposomes of defined C/L ratios. Subsequently, net 45Calcium-influx into the cell was measured by inhibiting the Ca-ejecting ATPase with vanadate. Additionally, the use of nitrendipine, a potent calcium channel inhibitor, during incubation allowed the determination of Ca-influx through the calcium channel. A positive correlation between the 45Ca++-influx and the molar C/L ratio of the membrane was found over a wide C/L range. A molar C/L ratio of 1.4 in the membrane increased calcium influx by 150 % compared to controls (molar C/L ratio = 0.8, calcium influx rate = 100 %), while a molar C/L ratio at less than 0.75 decreased calcium influx by 50 %. We conclude, that the cholesterol content of the membrane greatly influences the calcium channel and thus plays a pivotal role for the availability of calcium as a second messenger. These findings may provide a link between high plasma cholesterol and the development of atherosclerosis as well as enhanced platelet aggregability.
为了研究胆固醇含量对钙进入通道的影响,使用人类红细胞作为模型系统。通过用特定胆固醇与卵磷脂比例(C/L比例)的脂质体孵育细胞(15小时,25摄氏度),对细胞膜的胆固醇与卵磷脂比例(C/L比例)进行实验性改变。随后,通过用钒酸盐抑制钙排出ATP酶来测量进入细胞的净45钙流入量。此外,在孵育过程中使用强效钙通道抑制剂尼群地平,可以确定通过钙通道的钙流入量。在较宽的C/L范围内,发现45Ca++流入量与细胞膜的摩尔C/L比例之间存在正相关。与对照(摩尔C/L比例 = 0.8,钙流入率 = 100%)相比,细胞膜中摩尔C/L比例为1.4时,钙流入量增加了150%,而摩尔C/L比例小于0.75时,钙流入量减少了50%。我们得出结论,细胞膜的胆固醇含量极大地影响钙通道,因此在钙作为第二信使的可用性方面起着关键作用。这些发现可能为高血浆胆固醇与动脉粥样硬化的发展以及血小板聚集性增强之间提供联系。