Transport of Pregabalin Via L-Type Amino Acid Transporter 1 (SLC7A5) in Human Brain Capillary Endothelial Cell Line.
Transport of Pregabalin Via L-Type Amino Acid Transporter 1 (SLC7A5) in Human Brain Capillary Endothelial Cell Line.
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DOI：
10.1007/s11095-018-2532-0
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发表时间：
2018-10-29
影响因子：
3.7
通讯作者：
Tomi M
中科院分区：
文献类型：
--
作者：
Takahashi Y;Nishimura T;Higuchi K;Noguchi S;Tega Y;Kurosawa T;Deguchi Y;Tomi M
抗癫痫药物普瑞巴林尽管亲脂性低，但仍能通过血脑屏障(BBB)。本研究旨在探讨L类氨基酸转运体(LAT1/SLC7A5和LAT2/SLC7A8)对普瑞巴林摄取的影响。采用LC-MS/MS联用技术检测体外培养的人血脑屏障模型HEK293细胞和hCMEC/D3细胞对普瑞巴林的摄取。定量RT-PCR法检测hCMEC/D3细胞LAT1mRNA的表达。在HEK293细胞中过表达LAT1，而不是LAT2，显著增加了细胞对普瑞巴林的摄取，并且LAT1介导的摄取是饱和的，Km为0.288×10-6 mm。在1 mM普瑞巴林存在下，LAT1介导的氨基酸摄取被特异性地几乎完全抑制。HCMEC/D3细胞对普瑞巴林的摄取是非钠依赖的，饱和的(Km = 为0.854 mM)，并被1 mM的大氨基酸、1 mM的L抑制剂2-氨基双环-(2，2，1)庚烷-2-羧酸和10μ的LAT1选择性抑制剂JPH203强烈抑制。我们的结果表明，LAT1，而不是LAT2，识别普瑞巴林为底物。提示LAT1参与血脑屏障的前加巴林转运。本文的在线版本(10.1007/s11095-0182532-0)包含补充材料，可供授权用户使用。
The anti-epileptic drug pregabalin crosses the blood-brain barrier (BBB) in spite of its low lipophilicity. This study was performed to determine whether L-type amino acid transporters (LAT1/SLC7A5 and LAT2/SLC7A8) contribute to the uptake of pregabalin. Pregabalin uptake by LATs-transfected HEK293 cells or hCMEC/D3 cells, an in vitro human BBB model, was measured by LC-MS/MS analysis. Expression of LAT1 mRNA in hCMEC/D3 cells was determined by quantitative RT-PCR analysis. Overexpression of LAT1, but not LAT2, in HEK293 cells significantly increased the cellular uptake of pregabalin, and the LAT1-mediated uptake was saturable with a Km of 0.288 mM. LAT1-mediated amino acid uptake was inhibited specifically and almost completely in the presence of 1 mM pregabalin. The uptake of pregabalin by hCMEC/D3 cells was sodium-independent, saturable (Km = 0.854 mM), and strongly inhibited by large amino acids at 1 mM, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, a specific system L inhibitor, at 1 mM and by JPH203, a LAT1-selective inhibitor, at 10 μM. Pregabalin uptake in hCMEC/D3 cells was also decreased by 75% by the silencing of LAT1 gene using LAT1 siRNA. Our results indicate that LAT1, but not LAT2, recognizes pregabalin as a substrate. It is suggested that LAT1 mediates pregabalin transport at the BBB. The online version of this article (10.1007/s11095-018-2532-0) contains supplementary material, which is available to authorized users.
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影响因子：
4.7
作者：
Deitmer, JW;Bröer, A;Bröer, S
通讯作者：
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影响因子：
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