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培养的原代大鼠星形胶质细胞砷酸盐摄取的表征。

Characterization of arsenate uptake by cultured primary rat astrocytes.

机构信息

Center for Biomolecular Interactions Bremen, University of Bremen, PO. Box 330440, 28334, Bremen, Germany.

出版信息

Neurochem Res. 2013 Sep;38(9):1785-90. doi: 10.1007/s11064-013-1081-5. Epub 2013 Jun 1.

Abstract

Arsenate is known to be accumulated by cultured astrocytes and to stimulate astrocytic glutathione export, but the arsenate uptake into astrocytes has not been characterized so far. To address this topic, we have exposed primary rat astrocyte cultures to arsenate and determined the cellular arsenic content by atomic absorption spectroscopy. Viable astrocytes accumulated arsenate in a time- and concentration-dependent manner. Their cellular arsenic content increased almost proportional with time for up to 60 min after application of arsenate. Analysis of the concentration-dependent increase in the specific arsenic content of the cells after 30 min of arsenate exposure revealed that cultured astrocytes take up arsenate with saturable kinetics by a transport process that has apparent KM- and Vmax-values of 1.7 ± 0.2 mM and 28 ± 4 nmol/(mg protein × 30 min), respectively. Arsenate uptake in viable astrocytes was strongly inhibited by the presence of phosphate or by lowering the incubation temperature to 4 °C and was completely abolished in a sodium ion-free medium. These results strongly suggest that the saturable temperature-dependent arsenate uptake into astrocytes is mediated by a sodium-dependent phosphate cotransporter.

摘要

砷酸盐已被证实可在培养的星形胶质细胞中积累,并刺激星形胶质细胞谷胱甘肽的输出,但目前尚未对星形胶质细胞摄取砷酸盐的过程进行描述。为了解决这个问题,我们将原代大鼠星形胶质细胞培养物暴露于砷酸盐中,并通过原子吸收光谱法测定细胞内的砷含量。存活的星形胶质细胞以时间和浓度依赖的方式摄取砷酸盐。在砷酸盐作用 60 分钟内,其细胞内砷含量几乎与时间呈正比增加。对砷酸盐暴露 30 分钟后细胞内特定砷含量的浓度依赖性增加进行分析表明,培养的星形胶质细胞通过一种转运过程以饱和动力学摄取砷酸盐,该转运过程的表观 KM 和 Vmax 值分别为 1.7±0.2 mM 和 28±4 nmol/(mg 蛋白×30 分钟)。在有磷酸盐存在的情况下,或在 4°C 下降低孵育温度时,存活的星形胶质细胞中砷酸盐的摄取会受到强烈抑制,而在无钠离子的培养基中则完全被抑制。这些结果强烈表明,星形胶质细胞中依赖于温度的、饱和的、钠离子依赖的磷酸盐共转运体介导了砷酸盐的摄取。

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