PXR-ABC drug transporters/CYP-mediated ursolic acid transport and metabolism in vitro and vivo
PXR-ABC drug transporters/CYP-mediated ursolic acid transport and metabolism in vitro and vivo
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PXR-ABC药物转运蛋白/CYP介导的熊果酸体内外转运和代谢
DOI：
10.1002/ardp.202000082
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发表时间：
2020-07-06
影响因子：
5.1
通讯作者：
Li，Yuhua
中科院分区：
文献类型：
--
作者：
Wen，Jinhua;Zhou，Ying;Li，Yuhua
在转基因细胞模型中研究了熊果酸的转运动力学和代谢动力学。然后，研究了5-孕烯-3-β-20-酮-16α-Carnile(PCN)激活孕烷X受体(PXR)后，熊果酸在大鼠体内的药代动力学特征及组织中三磷酸腺苷结合盒转运体(ABC Transporter)和细胞色素P450(Cyp)酶的表达。在Caco 2-siRNA-PXR细胞中沉默PXR后，P-糖蛋白、乳腺癌耐药蛋白、多药耐药相关蛋白2(MRP2)和CYP2C9的蛋白丰度降低。10、20和50µM熊果酸在CaCo2细胞中的表观通透性分别为2.19 ± 0.44、1.40 ± 0.17和1.25 ± 0.07，而在CaCo2-siRNA-PxR细胞中分别为1.85 ± 0.36、1.24 ± 0.11和1.19 ± 0.04。PXR-RXRα可显著激活细胞内ABC转运蛋白的表达。与CACO2细胞相比，当熊果酸浓度为10、20和50 µM时，PxR激活后的CACO2-PxR-RxRα细胞的PDR值分别为1.60 ± 0.31比1.97 ± 0.21、1.46 ± 0.08比2.01 ± 0.19和1.32 ± 0.26比2.09 ± 0.22。同时，PXR-RXRα还能激活细胞色素P450代谢酶裂解产物中熊果酸的代谢动力学，Km值分别为81.99 ± 44.32和60.05 ± 29.62 /ml，Vmax分别为3.77 ± 0.86和3.41 ± 0.96 µg · ml−1· −1。细胞色素P3A4和细胞色素P3 4A的VmandKm值分别为3.57 ± 1.12 · m l−1· m in−1和81.71 ± 18.38 /m l，3.85 ± 1.46 m l · m l−1· m in−1和62.18 ± 14.56 m l.PCN作为小鼠PXR的强激动剂，可显著影响熊果酸在大鼠体内的药代动力学，对组织中CYP和转运蛋白的信使RNA表达有不同程度的影响。
The transporting kinetics and metabolic kinetics of ursolic acid were studied in transgenic cell models. Then, the pharmacokinetics features of ursolic acid and the expression of ATP‐binding cassette transporters (ABC transporter) and cytochrome P450 (CYP) enzymes in tissues after pregnane X receptor (PXR) activation by 5‐pregnen‐3β‐ol‐20‐one‐16α‐carbonitrile (PCN) were investigated in rats. After silencing of PXR in Caco2–siRNA–PXR cells, there was a decrease in the protein abundance of P‐glycoprotein, breast cancer‐resistant protein, multidrug resistance‐associated protein 2 (MRP2), and CYP2C9. The apparent permeability (PDR) values of 10, 20, and 50 µM ursolic acid in Caco2 cells were 2.19 ± 0.44, 1.40 ± 0.17, and 1.25 ± 0.07, respectively, whereas in Caco2–siRNA–PXR cells, they were 1.85 ± 0.36, 1.24 ± 0.11, and 1.19 ± 0.04, respectively. PXR–RXRα would significantly activate ABC transporter expression in Caco2 cells. Compared with Caco2 cells, when the concentrations of ursolic acid were 10, 20, and 50 µM, the PDR values increased in Caco2–PXR–RXRα cells after PXR activation: 1.60 ± 0.31 versus 1.97 ± 0.21, 1.46 ± 0.08 versus 2.01 ± 0.19, and 1.32 ± 0.26 versus 2.09 ± 0.22, respectively. Simultaneously, PXR–RXRα would activate the expression of CYP2C9; metabolic kinetics of ursolic acid in CYP metabolizing enzyme lysate of Caco2 cells and Caco2–PXR–RXR cells was studied and it was found that theKmvalues were 81.99 ± 44.32 and 60.05 ± 29.62 µg/ml, andVmaxvalues were 3.77 ± 0.86 and 3.41 ± 0.96 µg · ml−1· min−1, respectively. However, in human CYP metabolizing recombinase, we found that both CYP2C9 and CYP34A were involved in the metabolism of ursolic acid.VmandKmvalues for CYP3A4 and CYP2C9 were 3.57 ± 1.12 µg · ml−1· min−1and 81.71 ± 18.38 µg/ml, 3.85 ± 1.46 µg · ml−1· min−1and 62.18 ± 14.56 µg/ml, respectively. As a strong agonist for mouse pxr, PCN could significantly affect pharmacokinetics of ursolic acid in rats, and it showed discrepant effects on messenger RNA expression of cyp and transporters in tissues.