The potential role of human multidrug resistance protein 1 (MDR1) and multidrug resistance-associated protein 2 (MRP2) in the transport of Huperzine A in vitro
The potential role of human multidrug resistance protein 1 (MDR1) and multidrug resistance-associated protein 2 (MRP2) in the transport of Huperzine A in vitro
复制标题
人多药耐药蛋白1（MDR1）和多药耐药相关蛋白2（MRP2）在石杉碱甲体外转运中的潜在作用
DOI：
10.1080/00498254.2019.1623935
复制
发表时间：
2019-06-05
期刊：
影响因子：
1.8
通讯作者：
Zhang, Chunbo
中科院分区：
文献类型：
--
作者：
Fei, Ziyan;Hu, Mengyun;Zhang, Chunbo
摘要1.超过30%的癫痫患者存在抗药性。P-糖蛋白（P-gp）和MRP 2介导的抗癫痫药物转运在耐药性癫痫中起重要作用。石杉碱甲（Hup-A）是一种天然化合物，可能具有治疗癫痫和阿尔茨海默病等神经系统疾病的潜力。在这项研究中，我们研究了人P-gp和MRP 2是否转运Hup-A。2.使用转染人P-gp或MRP 2的LLC-PK 1和MDCK II细胞建立浓度平衡转运试验（CETA），并测定Hup-A的转运谱。采用qPCR和western blotting检测P-gp和MRP 2的表达。Rho 123和CDFDA细胞摄取法检测P-gp和MRP 2的转运功能。3.在CETA中，浓度为10 ng/mL或2 µg/mL的Hup-A通过MDR 1和MRP 2从细胞单层的基底侧转运至顶侧。P-gp和MRP 2抑制剂完全阻断Hup-A的外排。在LLC-PK 1或MDCKII野生型（WT）细胞中没有Hup-A的外排。4.我们证明Hup-A是P-gp和MRP 2的底物。这些结果暗示了Hup-A在体内穿过血脑屏障（BBB）的外排，表明Hup-A的潜在耐药性。
Abstract 1. More than 30% of epilepsy patients suffer pharmacoresistance. Transport of antileptic drugs by P-glycoprotein (P-gp) and MRP2 plays an important role in drug-resistant epilepsy. Huperzine A (Hup-A) is a natural compound, which might have potential in treating neurological disorders including epilepsy and Alzheimer’s disease. In this study, we investigated whether human P-gp and MRP2 transport Hup-A. 2. LLC-PK1 and MDCKII cells transfected with human P-gp or MRP2 were used to establish concentration equilibrium transport assays (CETAs) and determine the transport profile of Hup-A. The expression of P-gp and MRP2 was detected by qPCR and western blotting. The transport function of P-gp and MRP2 was measured by Rho123 and CDFDA cell uptake assay. 3. In CETAs, Hup-A at concentrations of 10 ng/mL or 2 µg/mL was transported by MDR1 and MRP2 from basolateral to apical sides of the cell monolayers. P-gp and MRP2 inhibitors completely blocked the efflux of Hup-A. There was no efflux of Hup-A in LLC-PK1 or MDCKII wild-type (WT) cells. 4. We demonstrate that Hup-A is a substrate of P-gp and MRP2. These results imply the efflux of Hup-A across the blood-brain barrier (BBB) in vivo, suggesting potential drug resistance of Hup-A.