Effect of polycyclic aromatic hydrocarbons on generation and efflux of glutathione conjugates in primary cultured alveolar epithelial cells.
Effect of polycyclic aromatic hydrocarbons on generation and efflux of glutathione conjugates in primary cultured alveolar epithelial cells.
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DOI：
10.2133/dmpk.19.407
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发表时间：
2004
影响因子：
2.1
通讯作者：
Keiko Nagayoshi;Takayuki Nemoto;Shumpei Yokoyama;F. Yamashita;M. Hashida
中科院分区：
文献类型：
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作者：
Keiko Nagayoshi;Takayuki Nemoto;Shumpei Yokoyama;F. Yamashita;M. Hashida
本研究旨在从功能上研究多药耐药相关蛋白(MRP)介导的跨肺上皮转运。从兔肺中分离出肺泡II型细胞，在Transwell上培养，直至形成致密的单层。在细胞单层中预加一氯双烷(MBCl)，并将其代谢成荧光谷胱甘肽结合物(mBCl-SG)后，定期测量输出到心尖室和基底室的mBCL-SG的量。MBCl-SG在根尖方向比在基底外侧更优先输出。MRP抑制剂MK-571可显著抑制肺泡上皮细胞mBCl-SG的外流。药代动力学分析表明，B[a]P增加mBCl-SG的外排不是由于MRP活性增强，而仅仅是由于细胞内mBCl-SG水平的增加。细胞内mBCl-SG水平的升高与苯并[a]P预处理引起的GSH降低有很好的对应关系。
This study was initiated to functionally characterize multidrug resistance associated protein (MRP)-mediated transport across the lung epithelium. Alveolar type II cells were isolated from rabbit lung and cultured on Transwell until forming a tight monolayers. After the cell monolayer was preloaded with monochlorobimane (mBCl) that is metabolized to a fluorescent glutathione conjugate (mBCl-SG), amount of mBCl-SG exported to apical and basal compartments were measured periodically. mBCl-SG was more preferentially exported in the apical direction than in the basolateral direction. Efflux of mBCl-SG from alveolar epithelial cells was significantly inhibited by a MRP inhibitor MK-571. Pharmacokinetic analysis of efflux profiles revealed that increased efflux of mBCl-SG by B[a]P is not due to enhanced MRP activity but simply due to an elevated level of mBCl-SG in the cells. Elevation of the intracellular level of mBCl-SG corresponded well to that of reduced GSH caused by B[a]P pretreatment.