Drug clearance from cerebrospinal fluid mediated by organic anion transporters 1 (Slc22a6) and 3 (Slc22a8) at arachnoid membrane of rats.
Drug clearance from cerebrospinal fluid mediated by organic anion transporters 1 (Slc22a6) and 3 (Slc22a8) at arachnoid membrane of rats.
复制标题
大鼠蛛网膜有机阴离子转运蛋白 1 (Slc22a6) 和 3 (Slc22a8) 介导的脑脊液药物清除。
DOI：
10.1021/acs.molpharmaceut.7b00852
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Terasaki T.
中科院分区：
文献类型：
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作者：
Zhang Z;Tachikawa M;Uchida Y;Terasaki T.
虽然蛛网膜上皮细胞形成血-蛛网膜屏障（BAB），充当血-CSF界面，但通常认为BAB对水溶性物质是不可渗透的，并且在很大程度上起被动作用。本研究旨在通过定量靶向绝对蛋白质组学（QTAP）和体内分析，阐明BAB转运蛋白在调节CSF清除水溶性有机阴离子药物中的作用。用QTAP法测定了大鼠软脑膜质膜组分中61种分子的蛋白表达水平，其中包括19种ATP结合盒（ABC）转运蛋白和32种溶质载体（SLC）转运蛋白。检测到33种蛋白质，其他蛋白质均低于定量限。多药耐药蛋白1（Mdr 1a/P-gp/Abcb 1a）和乳腺癌耐药蛋白（Bcrp/Abcg 2）的表达水平分别为16.6和3.27 fmol/μg蛋白（分别是脉络丛中的51.9倍和9.82倍）。在仅在软脑膜而非脉络丛检测到的有机阴离子转运蛋白中，有机阴离子转运蛋白1（oat 1/Slc 22 a6）显示最大表达（2.73 fmol/μg蛋白）。另一方面，Oat 3在软脑膜的蛋白表达水平为6.65 fmol/μg蛋白，与脉络丛的差异在2倍以内。为了研究燕麦1的作用，我们将对氨基马尿酸（PAH）与或不与燕麦1抑制剂注射到大鼠的小脑延髓池（以尽量减少脉络丛功能的贡献）。在长达15分钟的时间内，未从CSF中摄取大量流动标记物FITC-菊粉，而PAH的摄取清除率为26.5 μL/min。PAH摄取被3 mM头孢噻吩（抑制燕麦1和燕麦3）完全阻断，而17%的PAH摄取被0.2 mM头孢噻吩（选择性抑制燕麦3）抑制。这些结果表明，在BAB处的oat 1和oat 3提供了独立于脉络丛的来自CSF的有机阴离子药物的独特清除途径。
Although arachnoid mater epithelial cells form the blood–arachnoid barrier (BAB), acting as a blood–CSF interface, it has been generally considered that the BAB is impermeable to water-soluble substances and plays a largely passive role. Here, we aimed to clarify the function of transporters at the BAB in regulating CSF clearance of water-soluble organic anion drugs based on quantitative targeted absolute proteomics (QTAP) andin vivoanalyses. Protein expression levels of 61 molecules, including 19 ATP-binding-cassette (ABC) transporters and 32 solute-carrier (SLC) transporters, were measured in plasma membrane fraction of rat leptomeninges using QTAP. Thirty-three proteins were detected; others were under the quantification limits. Expression levels of multidrug resistance protein 1 (Mdr1a/P-gp/Abcb1a) and breast cancer resistance protein (Bcrp/Abcg2) were 16.6 and 3.27 fmol/μg protein (51.9- and 9.82-fold greater than in choroid plexus, respectively). Among those organic anion transporters detected only at leptomeninges, not choroid plexus, organic anion transporter 1 (oat1/Slc22a6) showed the greatest expression (2.73 fmol/μg protein). On the other hand, the protein expression level of oat3 at leptomeninges was 6.65 fmol/μg protein, and the difference from choroid plexus was within two-fold. To investigate oat1’s role, we injected para-aminohippuric acid (PAH) with or without oat1 inhibitors into cisterna magna (to minimize the contribution of choroid plexus function) of rats. A bulk flow marker, FITC-inulin, was not taken up from CSF up to 15 min, whereas uptake clearance of PAH was 26.5 μL/min. PAH uptake was completely blocked by 3 mM cephalothin (inhibits both oat1 and oat3), while 17% of PAH uptake was inhibited by 0.2 mM cephalothin (selectively inhibits oat3). These results indicate that oat1 and oat3 at the BAB provide a distinct clearance pathway of organic anion drugs from CSF independently of choroid plexus.