Institute for Innovative Drug Design and Evaluation, School of Pharmacy, Henan University, N. Jinming Ave., Kaifeng, Henan 475004, China.
Institute for Innovative Drug Design and Evaluation, School of Pharmacy, Henan University, N. Jinming Ave., Kaifeng, Henan 475004, China.
Eur J Pharm Sci. 2018 Nov 1;124:228-239. doi: 10.1016/j.ejps.2018.08.041. Epub 2018 Aug 31.
This study aimed to investigate the cellular disposition of liquiritigenin via the sulfonation pathway and the role of efflux transporters in liquiritigenin sulfate excretion. The sulfonation disposition of liquiritigenin was investigated using SULT1A3 overexpressed HEK293 cells (HEK-SULT1A3 cells). Liquiritigenin generated one mono-sulfate metabolite (7-O-sulfate) in HEK-SULT1A3 cell lysate. And the sulfonation followed the Michaelis-Menten kinetic (V = 0.84 nmol/min/mg and K = 7.12 μM). Expectedly, recombinant SULT1A3 (hSULT1A3) showed a highly similar kinetic profile with cell lysate. Furthermore, 7-O-sulfate was rapidly generated and excreted in HEK-SULT1A3 cells. Ko143 (a BCRP-selective inhibitor) at 20 μM significantly decreased the excretion rate of liquiritigenin sulfate (>42.5%, p < 0.001). Moreover, the pan-MRPs inhibitor MK-571 at 20 μM essentially abolished the liquiritigenin sulfate effluxion, resulting in the marked reduction of excretion rate (>97.4%, p < 0.001). Furthermore, knockdown of BCRP led to moderate reduction in sulfate excretion (15.9%-16.9%, p < 0.05). Silencing of MRP4 caused significant decreased in sulfate excretion (20.2%-32.5%, p < 0.01). In conclusion, one sulfate metabolite was generated from liquiritigenin in HEK-SULT1A3 cells. BCRP and MRP4 should be the key factors for the cellular excretion of liquiritigenin sulfate.
本研究旨在通过磺化途径研究甘草素的细胞分布，以及外排转运体在甘草素硫酸排泄中的作用。使用过表达 SULT1A3 的 HEK293 细胞（HEK-SULT1A3 细胞）研究甘草素的磺化分布。在 HEK-SULT1A3 细胞裂解液中，甘草素生成一种单硫酸盐代谢物（7-O-硫酸盐）。磺化反应符合米氏动力学（V=0.84 nmol/min/mg，K=7.12 μM）。不出所料，重组 SULT1A3（hSULT1A3）与细胞裂解液表现出高度相似的动力学特征。此外，7-O-硫酸盐在 HEK-SULT1A3 细胞中迅速生成并排泄。20 μM 的 Ko143（BCRP 选择性抑制剂）显著降低甘草素硫酸的排泄率（>42.5%，p<0.001）。此外，20 μM 的泛 MRPs 抑制剂 MK-571 基本上消除了甘草素硫酸盐的外排作用，导致排泄率显著降低（>97.4%，p<0.001）。此外，BCRP 的敲低导致硫酸盐排泄适度减少（15.9%-16.9%，p<0.05）。MRP4 的沉默导致硫酸盐排泄显著减少（20.2%-32.5%，p<0.01）。综上所述，在 HEK-SULT1A3 细胞中，甘草素生成一种硫酸盐代谢物。BCRP 和 MRP4 可能是甘草素硫酸细胞排泄的关键因素。
