The First Affiliated Hospital of Jinzhou Medical University.
Jiangxi University of Traditional Chinese Medicine.
Chem Pharm Bull (Tokyo). 2022;70(10):669-678. doi: 10.1248/cpb.c22-00093.
This study aimed to evaluate the interspecies difference in metabolism of mulberrin and examine the interaction between mulberrin and CYP enzymes or recombinant human uridine 5'-diphosphate (UDP)-glucuronosyltransferase (UGT) enzymes. Liver microsomes from human (HLMs), Beagle dog (DLMs), minipig (PLMs), monkey (MLMs), rabbit (RLMs), rat (RAMs), and mouse (MIMs) were used to investigate metabolic diversity among different species. Additionally, recombinant human supersomes were used to confirm that metabolic enzymes are involved in the biotransformation of mulberrin. We also evaluated the influence of mulberrin on protein expression by Western blot analysis. Mulberrin metabolism showed significant interspecies differences. We found four and two metabolites in phase I and II reaction systems, respectively. In phase I metabolism profiles of mulberrin for HLMs, PLMs and MLMs conformed to the classic Michaelis-Menten kinetics, RAMs and MIMs followed biphasic kinetics; phase II reaction of mulberrin in HLMs, DLMs, PLMs, MLMs, RLMs, RAMs and MIMs followed biphasic kinetics. UGT1A1 were the major CYP isoforms responsible for the metabolism of mulberrin. Mulberrin showed potent inhibitory effects against CYP3A4, CYP2C9, CYP2E1, UGT1A1, UGT1A3 and UGT2B7 with IC values of 54.21, 9.93, 39.12, 3.84, 2.01, 16.36 µM, respectively. According to Western blot analysis, mulberrin can upregulate the protein expression of CYP2C19, and downregulate the expression levels of CYP3A5 and CYP2C9 in HepG2 cells as concentration increased. The interspecies comparisons can help find other species with metabolic pathways similar to those in humans for future in vivo studies.
本研究旨在评估桑枝木聚糖在不同物种间的代谢差异，并研究桑枝木聚糖与 CYP 酶或重组人尿苷 5'-二磷酸（UDP）-葡萄糖醛酸基转移酶（UGT）酶之间的相互作用。用人（HLMs）、比格犬（DLMs）、小型猪（PLMs）、猴（MLMs）、兔（RLMs）、大鼠（RAMs）和小鼠（MIMs）的肝微粒体来研究不同物种间的代谢多样性。此外，还使用重组人超微粒体来确认代谢酶参与了桑枝木聚糖的生物转化。我们还通过 Western blot 分析评估了桑枝木聚糖对蛋白质表达的影响。桑枝木聚糖的代谢在不同物种间存在显著差异。我们在 I 相和 II 相反应系统中分别发现了 4 种和 2 种代谢产物。在 HLMs、PLMs 和 MLMs 的桑枝木聚糖 I 相代谢谱中，符合经典的米氏动力学；而在 RAMs 和 MIMs 中则遵循双相动力学；HLMs、DLMs、PLMs、MLMs、RLMs、RAMs 和 MIMs 的桑枝木聚糖 II 相反应遵循双相动力学。UGT1A1 是负责桑枝木聚糖代谢的主要 CYP 同工酶。桑枝木聚糖对 CYP3A4、CYP2C9、CYP2E1、UGT1A1、UGT1A3 和 UGT2B7 具有很强的抑制作用，IC 值分别为 54.21、9.93、39.12、3.84、2.01、16.36µM。根据 Western blot 分析，桑枝木聚糖可以上调 HepG2 细胞中 CYP2C19 的蛋白表达水平，并随着浓度的增加下调 CYP3A5 和 CYP2C9 的表达水平。种间比较有助于找到与人类代谢途径相似的其他物种，用于未来的体内研究。
