L-type Amino Acid Transporter 1 (SLC7A5)-Mediated Transport of Pregabalin at the Rat Blood-Spinal Cord Barrier and its Sensitivity to Plasma Branched-Chain Amino Acids
L-type Amino Acid Transporter 1 (SLC7A5)-Mediated Transport of Pregabalin at the Rat Blood-Spinal Cord Barrier and its Sensitivity to Plasma Branched-Chain Amino Acids
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DOI：
10.1016/j.xphs.2022.12.028
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发表时间：
2023-03-16
影响因子：
3.8
通讯作者：
Tomi, Masatoshi
中科院分区：
文献类型：
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作者：
Akashi, Tomoya;Noguchi, Saki;Tomi, Masatoshi
普瑞巴林是一种抗神经性疼痛药物，可抑制脊髓中电压依赖性钙通道的a28亚单位。本研究的目的是通过大鼠体内实验和原代培养的大鼠脊髓内皮细胞的体外研究来表征普瑞巴林在血脊髓屏障（BSCB）的转运机制。我们通过在嘌呤霉素存在下培养大鼠脊髓组织来分离内皮细胞，并证实了BSCB标记物如Cd 31、Mdr 1a和Claudin-5的表达。原代培养的大鼠脊髓内皮细胞对普瑞巴林的摄取是钠非依赖性的，并且被L-亮氨酸、2-氨基二环-（2，2，1）-庚烷-2-羧酸和JPH 203显著抑制。这些结果表明L型氨基酸转运蛋白（LAT）1的参与。在原代培养的大鼠脊髓内皮细胞中表达LAT 1 mRNA和蛋白，这与BSCB中的LAT 1表达一致。在体内研究中，通过预先给药支链氨基酸（BCAAs）（LAT 1的内源性底物），普瑞巴林向大鼠脊髓和脑的转移显著减少。我们的研究结果表明，普瑞巴林跨BSCB的转运至少部分由LAT 1介导，并被血浆BCAAs抑制。(c)2023年由Elsevier Inc.出版代表美国药学协会
Pregabalin is an anti-neuropathic pain drug inhibiting the a28 subunit of the voltage-dependent calcium channel in the spinal cord. The aim of this study is to characterize the transport mechanism of pregabalin at the blood-spinal cord barrier (BSCB) by means of in vivo experiments in rats and in vitro studies using primary-cultured rat spinal cord endothelial cells. We isolated endothelial cells by culturing rat spinal cord tissue in the presence of puromycin, and confirmed the expression of BSCB markers such as Cd31, Mdr1a, and Claudin-5. The uptake of pregabalin by primary-cultured rat spinal cord endothelial cells was sodium-independent and was significantly inhibited by L-leucine, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, and JPH203. These results suggest the involvement of L-type amino acid transporter (LAT) 1. LAT1 mRNA and protein was expressed in primary-cultured rat spinal cord endothelial cells, which is consistent with LAT1 expression at the BSCB. In the in vivo study, the transfer of pregabalin to rat spinal cord and brain was significantly decreased by the pre-administration of branched chain amino acids (BCAAs), which are endogenous substrates of LAT1. Our results indicate that pregabalin transport across the BSCB is mediated at least in part by LAT1 and is inhibited by plasma BCAAs.(c) 2023 Published by Elsevier Inc. on behalf of American Pharmacists Association.