Use of rhodamine 123 to examine the functional activity of P-glycoprotein in primary cultured brain microvessel endothelial cell monolayers
Use of rhodamine 123 to examine the functional activity of P-glycoprotein in primary cultured brain microvessel endothelial cell monolayers
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DOI：
10.1016/0024-3205(96)00483-3
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发表时间：
1996-09-27
期刊：
影响因子：
6.1
通讯作者：
Miller, DW
中科院分区：
文献类型：
--
作者：
Fontaine, M;Elmquist, WF;Miller, DW
用荧光染料罗丹明123检测了原代培养的牛脑微血管内皮细胞（BBMEC）单层P-糖蛋白（P-gp）外排转运系统的功能活性。罗丹明123的积累与P-gp修饰剂，环孢素A（CSA）处理的BBMEC单层显着增加。罗丹明123的积累也增加了其他P-gp修饰剂。这些药物增加罗丹明123在BBMEC单层中蓄积的等级有效性为CSA=双嘧达莫>维拉帕米=奎尼丁。CSA处理的BBMEC单层中罗丹明123蓄积的最大增加约为对照单层的3倍，定性上与3 H-长春新碱观察到的结果相似。功能活性与BBMEC单层和过表达P-gp的既定肿瘤细胞系中P-gp的生化表达的比较表明，功能活性可能是比蛋白质表达更能描述该药物外排系统重要性的指标。此外，这些研究表明，罗丹明123在BBMEC单层中的积累可用于定量检测血脑屏障中的P-gp活性。
The fluorescent dye, rhodamine 123, was used to evaluate the functional activity of the P-glycoprotein (P-gp) efflux transport system in primary cultured bovine brain microvessel endothelial cell (BBMEC) monolayers. Rhodamine 123 accumulation was increased significantly in BBMEC monolayers treated with the P-gp modifying agent, cyclosporin A (CSA). Rhodamine 123 accumulation was also increased by other P-gp modifying agents. The rank effectiveness of these agents in increasing rhodamine 123 accumulation in BBMEC monolayers was CSA=dipyridamole>verapamil=quinidine. The maximal increase in rhodamine 123 accumulation in CSA treated BBMEC monolayers was approximately 3 fold greater than in control monolayers and was qualitatively similar to that observed with 3H-vincristine. Comparison of functional activity with the biochemical expression of P-gp in BBMEC monolayers and in an established tumor cell line that over-expresses P-gp indicate that functional activity may be a more descriptive measure of the importance of this drug efflux system than protein expression. Furthermore, these studies suggest that accumulation of rhodamine 123 in BBMEC monolayers can be used to quantitatively examine P-gp activity in the blood-brain barrier.