齐墩果酸在人及其他种属肝微粒体CYP450中代谢的差异研究

摘要

Objective To compare the metabolism of oleanolic acid in liver microsomes of human and other species.Methods An ultra-high performance liquid chromatography/mass spectrometry (UPLC/MS) method was applied to systemically study the oleanolic acid metabolism in liver microsomes of human, rat, mouse, pig, rabbit, cattle, and sheep, and comprehensively evaluate oleanolic acid metabolites in vitro and metabolism differences between species.The research also tested the metabolites after incubation with selective inhibitors of 4 main P450s including CYP2C9, CYP2D6, CYP2E1, and CYP3A4 in liver microsomes.Results The results showed that the most metabolite M-469 was generated in Kunming mouse liver microsomes, followed by C57 mouse, SD rat, human, pig, and rabbit.However, in the cattle and sheep liver microsome incubation system, the metabolite M-469 was not found.For human, CYP3A4 was identified as the main enzyme responsible for the metabolite M-469 of oleanolic acid.For Kunming mouse, CYP3A and CYP2E1 were involved in the production of this metabolite.Conclusion There was significant species difference in oleanolic acid metabolism in vitro between human and other species, providing important reference value for the further metabolism study and safety evaluation of oleanolic acid.%目的 比较齐墩果酸在人和其他种属肝微粒体CYP450酶中的代谢差异.方法 采用基于多重质量亏损(Multiple mass defect filter, MMDF)的四极杆-静电场轨道阱高分辨质谱色谱联用仪,分析齐墩果酸经人及昆明小鼠、C57小鼠、SD大鼠、猪、牛、羊、兔肝微粒体体外代谢的产物及种属差异.利用与CYP450各亚型酶的特异性抑制剂共孵育,对比不同种属参与齐墩果酸特征性代谢产物M-469的主要代谢亚型酶.结果 齐墩果酸不同种属的肝微粒体体外代谢存在明显差异.齐墩果酸特征性代谢产物M-469,在昆明小鼠的肝微粒体中生成最多,其次是C57小鼠、SD大鼠、人、猪、兔,而牛、羊体外不生成此产物.在人肝微粒体CYP450酶中,CYP3A4是齐墩果酸特征性代谢产物M-469的主要代谢酶.昆明小鼠CYP450酶中,CYP3A、CYP2E1参与了此代谢产物的生成.结论 齐墩果酸在人和不同种属肝微粒体CYP450代谢中存在差异,因此,本实验将为齐墩果酸进一步代谢研究及安全性评价提供重要参考价值.