Khodorova A B, Astashkin E I
Institute of Biotechnology, Moscow, Russian Federation.
FEBS Lett. 1994 Oct 17;353(2):167-70. doi: 10.1016/0014-5793(94)01029-3.
A sustained increase of cytoplasmic Ca2+ concentration ([Ca2+]i) in lymphocytes was induced by a potent inhibitor of the intracellular Ca(2+)-pump 2,5-di-(tert-butyl)-1,4-hydroquinone (BHQ) that effectively depletes Ca2+ stores and produces Ca2+ influx. The addition of low concentrations of arachidonic acid (AA) (less than 10 microM) in the presence of a BHQ-induced Ca2+ response produced a [Ca2+]i decrease. At the same time at higher concentrations, AA induced an additional increase of [Ca2+]i. The unidirectional uptake of Mn2+ into fura-2 loaded thymocytes was used to examine the effects of AA and BHQ on Ca2+ permeability of the plasma membrane. Judging by the rate of Mn(2+)-induced fura-2 fluorescence quenching, AA causes a concentration-dependent increase of the divalent cations' permeability both in the control solution and in the presence of BHQ. These results allowed us to suggest that the decrease of BHQ-induced [Ca2+]i stationary levels caused by low AA concentrations is due to the enhancement of Ca2+ extrusion from the cell, whereas the additional [Ca2+]i increase at high AA concentrations results from an elevation of cell membrane Ca2+ permeability.
细胞内钙泵的强效抑制剂2,5 - 二 -(叔丁基)-1,4 - 对苯二酚(BHQ)可诱导淋巴细胞胞质钙离子浓度([Ca2+]i)持续升高,该抑制剂能有效耗尽钙离子储备并促使钙离子内流。在BHQ诱导的钙离子反应存在的情况下,添加低浓度花生四烯酸(AA)(小于10微摩尔)会使[Ca2+]i降低。同时,在较高浓度时,AA会使[Ca2+]i进一步升高。利用Mn2+单向摄取到负载fura - 2的胸腺细胞中的方法,来检测AA和BHQ对质膜钙离子通透性的影响。根据Mn(2+)诱导的fura - 2荧光淬灭速率判断,AA在对照溶液以及存在BHQ的情况下,都会导致二价阳离子通透性呈浓度依赖性增加。这些结果使我们推测,低浓度AA导致BHQ诱导的[Ca2+]i稳定水平降低,是由于细胞内钙离子外排增强所致,而高浓度AA时[Ca2+]i的额外升高是由于细胞膜钙离子通透性增加所致。