Xenobiotic transport across isolated brain microvessels studied by confocal microscopy
Xenobiotic transport across isolated brain microvessels studied by confocal microscopy
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DOI：
10.1124/mol.58.6.1357
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发表时间：
2000-12-01
影响因子：
3.6
通讯作者：
Fricker, G
中科院分区：
文献类型：
--
作者：
Miller, DS;Nobmann, SN;Fricker, G
为了鉴定将外源性物质从中枢神经系统驱动到血液的特定转运蛋白，使用共聚焦显微镜和定量图像分析研究了荧光药物在大鼠和猪脑的分离毛细血管中的积累。道诺霉素和环孢素A（CSA）和伊维菌素的荧光衍生物的管腔积累是集中的，具体的，和能量依赖性（抑制NaCN）。PSC 833、伊维菌素、维拉帕米、CSA和钒酸盐降低了转运，但白三烯C-4（LTC 4）未降低转运，表明P-糖蛋白参与。荧光有机阴离子磺酰罗丹明101和荧光素甲氨蝶呤的管腔积累也是浓度，特异性和能量依赖性的。LTC 4、氯二硝基苯和钒酸盐减少了这些化合物的转运，但PSC 833和维拉帕米没有，表明涉及多药耐药相关蛋白（Mrp）。免疫组化定位的P-糖蛋白和Mrp 2的毛细血管内皮细胞和定量聚合酶链反应显示Mrp 1和Mrp 2的表达腔表面。最后，HIV蛋白酶抑制剂沙奎那韦和利托那韦是P-糖蛋白和Mrp介导的转运的有效抑制剂。这些结果验证了一种新的方法，用于研究药物转运在分离的脑毛细血管和牵连P-糖蛋白和一个或多个成员的Mrp家族的药物转运从中枢神经系统的血液。
To identify specific transporters that drive xenobiotics from central nervous system to blood, the accumulation of fluorescent drugs was studied in isolated capillaries from rat and pig brain using confocal microscopy and quantitative image analysis. Luminal accumulation of daunomycin and of fluorescent derivatives of cyclosporine A (CSA) and ivermectin was concentrative, specific, and energy-dependent (inhibition by NaCN). Transport was reduced by PSC 833, ivermectin, verapamil, CSA, and vanadate, but not by leukotriene C-4 (LTC4), indicating the involvement of P-glycoprotein. Luminal accumulation of the fluorescent organic anions sulforhodamine 101 and fluorescein methotrexate was also concentrative, specific, and energy-dependent. LTC4, chlorodinitrobenzene, and vanadate reduced transport of these compounds, but PSC 833 and verapamil did not, indicating the involvement of a multidrug resistance-associated protein (Mrp). Immunostaining localized P-glycoprotein and Mrp2 to the luminal surface of the capillary endothelium and quantitative polymerase chain reaction showed Mrp1 and Mrp2 expression. Finally, the HIV protease inhibitors saquinavir and ritonavir were potent inhibitors of transport mediated by both P-glycoprotein and Mrp. These results validate a new method for studying drug transport in isolated brain capillaries and implicate both P-glycoprotein and one or more members of the Mrp family in drug transport from central nervous system to blood.