Isolation of endothelial cells from brain, lung, and kidney:: Expression of the multidrug resistance P-glycoprotein isoforms
Isolation of endothelial cells from brain, lung, and kidney:: Expression of the multidrug resistance P-glycoprotein isoforms
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DOI：
10.1006/bbrc.2001.4312
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发表时间：
2001-03-02
影响因子：
3.1
通讯作者：
Béliveau, R
中科院分区：
文献类型：
--
作者：
Demeule, M;Labelle, M;Béliveau, R
使用磁性微珠与针对血小板-内皮细胞黏附分子-1(PECAM-1)的抗体交联，从脑、肺和肾皮质分离内皮细胞(EC)。用免疫印迹法检测内皮型一氧化氮合酶(ENOS)和PECAM-1的水平，两者都在阳性选择的EC组分中得到丰富。多药耐药P-糖蛋白(P-gp)在脑EC组分中高度浓缩(59倍)，而在阴性组分中缺失，其中存在星形胶质细胞标志物胶质纤维酸性蛋白(GFAP)。EC组肾皮质和肺组织P-gp水平较低。逆转录-聚合酶链式反应分析表明，mdr1a基因在脑组织EC组分中优先表达。Mdr1b基因在肾皮质EC中表达，而两种mdr1基因均在肺EC中表达。我们的结果表明，可以用微球分离EC，在脑中发现的P-gp的亚型主要是与EC相关的mdr1a。(C)2001年学术出版社。
Endothelial cells (EC) were isolated from brain, lung, and renal cortex using magnetic microbeads cross-linked to an antibody directed against the platelet-endothelial cell adhesion molecule-1 (PECAM-1). Levels of endothelial nitric oxide synthase (eNOS) and PECAM-1 were measured by Western blots and both were enriched in the positively selected EC fractions. The multidrug resistance P-glycoprotein (P-gp) was strongly enriched (59-fold) in the EC fraction from brain and was absent in the negative fraction, in which the glial fibrillary acidic protein (GFAP), an astrocyte marker, was present. Lower P-gp levels were detected in EC from renal cortex and lung. Reverse transcription-polymerase chain reaction analysis showed that the mdr1a gene was preferentially expressed in EC fraction from the brain. The mdr1b gene was found in EC from renal cortex whereas both mdr1 genes were detected in EC from lung. Our results indicate that EC can be isolated using microbeads and that the isoform of P-gp found in brain is mostly mdr1a, associated with EC. (C) 2001 Academic Press.