Endothelin-1 Reduces P-Glycoprotein Transport Activity in an In Vitro Model of Human Adult Blood-brain Barrier
Endothelin-1 Reduces P-Glycoprotein Transport Activity in an In Vitro Model of Human Adult Blood-brain Barrier
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DOI：
10.1007/s10571-008-9277-y
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发表时间：
2008-11-01
影响因子：
4
通讯作者：
Mabondzo, Aloise
中科院分区：
文献类型：
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作者：
Hembury, Alexandra;Mabondzo, Aloise
目的 了解血脑屏障 (BBB) 是一个巨大的挑战，它是与许多脑部疾病相关的神经炎症的关键因素。 BBB 还调节异生物质进入中枢神经系统 (CNS) 的通道，从而影响药物疗效。这可能是由于 BBB 上存在 ATP 结合盒转运蛋白，例如 P-糖蛋白 (Pgp)，它们是已知可转运许多药物的外排泵。肽内皮素 1 (ET-1) 与多种 CNS 疾病和神经炎症有关，并且已知可调节 Pgp 转运活性。尽管有动物模型的数据，但人体模型的数据却很少。当将成人体外 BBB 模型暴露于 ET-1 时，我们评估了成人大脑微血管内皮细胞 (HBMEC) 中 Pgp 的表达和转运活性。方法 成人 HBMEC 与人成人神经胶质细胞在 Transwells(R) R 上共培养，以模拟血液和中枢神经系统隔室。这些人类体外 BBB 暴露于 100 nM 和 10 nM ET-1 24 小时。通过流式细胞术评估 Pgp 表达，并通过测量放射性标记的地高辛通道评估其转运活性。结果暴露于 ET-1 后，流式细胞术显示 Pgp 特异性抗体的荧光强度没有变化。对于 10 nM ET-1，地高辛的通过增加，Q 比显着降低。结论 我们的结果表明，ET-1 对成人 HBMEC 的 Pgp 表达没有影响，但确实调节 Pgp 转运活性。
Aims It is a huge challenge to understand the blood-brain barrier (BBB), which is a key element in neuroinflammation associated with many brain diseases. The BBB also regulates the passage of xenobiotics into the central nervous system (CNS), and therefore influences drug efficacy. This may be due to the presence of ATP binding cassette transporters such as P-glycoprotein (Pgp) on the BBB, which are efflux pumps known to transport many drugs. The peptide endothelin 1 ( ET-1) is involved in different kinds of CNS diseases and neuroinflammation, and is known to modulate Pgp transport activity. Although there are data from animal models, data from human models are scarce. We evaluated Pgp expression and transport activity in adult human brain microvascular endothelial cells (HBMECs) when exposing an adult human in vitro BBB model to ET-1. Methods Adult HBMECs were cocultured with human adult glial cells on a Transwells(R) R to mimic blood and CNS compartments. These human in vitro BBBs were exposed for 24 h to 100 nM and 10 nM ET-1. Pgp expression was assessed by flow cytometry and its transport activity by measuring radiolabelled digoxin passage. Results After exposure to ET-1, flow cytometry showed no shift of fluorescence intensity for a Pgp specific antibody. The passage of digoxin increased with a significant decrease of Q ratio for 10 nM ET-1. Conclusion Our results show that ET-1 has no effect on Pgp expression of adult HBMECs, but does modulate Pgp transport activity.