Interplay of Efflux Transporters with Glucuronidation and Its Impact on Subcellular Aglycone and Glucuronide Disposition: A Case Study with Kaempferol.
Interplay of Efflux Transporters with Glucuronidation and Its Impact on Subcellular Aglycone and Glucuronide Disposition: A Case Study with Kaempferol.
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DOI：
10.1021/acs.molpharmaceut.8b00782
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发表时间：
2018-10
影响因子：
4.9
通讯作者：
Yuhuan Li;Linlin Lu;Liping Wang;W. Qu;Wenqin Liu;Yushan Xie;Hongming Zheng;Ying Wang;Xiao-xiao Qi;Ming Hu;Lijun J. Zhu;Zhongqiu Liu
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文献类型：
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作者：
Yuhuan Li;Linlin Lu;Liping Wang;W. Qu;Wenqin Liu;Yushan Xie;Hongming Zheng;Ying Wang;Xiao-xiao Qi;Ming Hu;Lijun J. Zhu;Zhongqiu Liu
葡萄糖醛酸化是药物代谢和消除的主要过程，通常控制药物疗效和毒性。出版物已证明外排转运蛋白控制细胞内葡萄糖醛酸化代谢。然而，目前还不清楚是否以及如何外排转运UDP-葡萄糖醛酸转移酶（UGT）在亚细胞器中相互作用。在这项研究中，山奈酚，一种模型荧光类黄酮，被用来研究葡萄糖醛酸化与运输在亚细胞水平上的相互作用。利用人重组UGT和微粒体表征山萘酚的体外葡萄糖醛酸化动力学。在Caco-2/TC 7细胞中，目视和定量测定UGT和外排转运蛋白对山奈酚亚细胞处置的抑制。视觉上评估了转运蛋白的敲除对山奈酚在肝和肠中的亚细胞积累的影响。通过活性氧（ROS）和核转录因子2（Nrf 2）的测定，评价山奈酚的药理活性。结果表明，UGT 1A 9是山奈酚葡萄糖醛酸化的主要酶。视觉和定量数据显示，UGT 1A 9抑制剂香芹酚导致山柰酚亚细胞苷元显着增加和亚细胞葡萄糖醛酸苷减少。抑制和敲除转运蛋白，如P-糖蛋白（P-gp），乳腺癌耐药蛋白（BCRP），和多药耐药相关蛋白（MRPs），表现出显着增加亚细胞山奈酚和减少其亚细胞葡萄糖醛酸苷。相应地，UGT 1A 9和转运蛋白的抑制导致山奈酚增加，因此，显著增强ROS清除效率和Nrf 2的核转位。总之，外排转运蛋白（P-gp、BCRP和MRP）和UGT的相互作用决定了山奈酚的亚细胞暴露和相应的药理学活性。
Glucuronidation is a major process of drug metabolism and elimination that generally governs drug efficacy and toxicity. Publications have demonstrated that efflux transporters control intracellular glucuronidation metabolism. However, it is still unclear whether and how efflux transporters interact with UDP-glucuronosyltransferases (UGTs) in subcellular organelles. In this study, kaempferol, a model fluorescent flavonoid, was used to investigate the interplay of glucuronidation with transport at the subcellular level. Human recombinant UGTs and microsomes were utilized to characterize the in vitro glucuronidation kinetics of kaempferol. The inhibition of UGTs and efflux transporters on the subcellular disposition of kaempferol were determined visually and quantitatively in Caco-2/TC7 cells. The knockout of transporters on the subcellular accumulation of kaempferol in liver and intestine were evaluated visually. ROS and Nrf2 were assayed to evaluate the pharmacological activities of kaempferol. The results showed that UGT1A9 is the primary enzyme responsible for kaempferol glucuronidation. Visual and quantitative data showed that the UGT1A9 inhibitor carvacrol caused a significant rise in subcellular aglycone and reduction in subcellular glucuronides of kaempferol. The inhibition and knockout of transporters, such as P-glycoprotein (P-gp), breast cancer resistance protein (BCRP), and multidrug resistance-associated proteins (MRPs), exhibited a marked increase in subcellular kaempferol and decrease in its subcellular glucuronides. Correspondingly, inhibition of UGT1A9 and transporters led to increased kaempferol and, consequently, a significantly enhanced ROS scavenging efficiency and nuclear translocation of Nrf2. In conclusion, the interplay of efflux transporters (P-gp, BCRP, and MRPs) and UGTs govern the subcellular exposure and corresponding pharmacological activity of kaempferol.