摘要
In the present study,we aimed to investigate the interaction between atractylenolideⅡ(AT-Ⅱ)and CYP450 enzyme in human liver microsomes,and to lay a theoretical foundation for predicting the possible interaction of AT-Ⅱin combination with drugs.The chemical inhibition experiment was carried out with specific inhibitors to clarify the CYP450 subtypes affecting the metabolism of AT-Ⅱ,and the mechanism,kinetics,and type of inhibition of CYP450 enzyme by AT-Ⅱwere studied by using the probe-based determination method of human liver microsome system with the related data of IC50 and Ki as evaluation indexes.The metabolism of AT-Ⅱwas affected by CYP1A2,CYP2C9 and CYP3A4 inhibitors,and the highest inhibition rates were41.35%,41.97%and 82.45%,respectively.The IC50 values of AT-Ⅱto five subtypes of P450 CYP2C9,CYP1A2,CYP2C19,CYP3A4 and CYP2D6 were 69.7,84.3,92.4,173.8 and 190.1μmol/L,respectively.The Ki values of AT-Ⅱto five subtypes of P450 CYP2C9,CYP1A2,CYP2C19,CYP3A4 and CYP2D6 were 190.6,179.1,>200,72.2 and 66.8,respectively.Among these enzymes,AT-Ⅱexhibited non-competitive inhibition on CYP1A2,showed competitive inhibition on CYP2C9 and CYP3A4,and displayed mixed AT-Ⅱinhibition on CYP2C19 and CYP2D6.CYP1A2,CYP2C9 and CYP3A4 were involved in the AT-Ⅱmetabolism,and AT-Ⅱexhibited different inhibitory mechanisms and strengths for the five subtypes of CYP450.
本研究旨在探讨白术内酯Ⅱ在人肝微粒体中与CYP450酶之间的相互作用情况,为预测白术内酯Ⅱ在药物联用时可能产生的相互作用奠定理论基础。采用特异性抑制剂进行化学抑制实验,阐明影响白术内酯Ⅱ代谢的CYP450亚型;以IC50和Ki等相关数据为评价指标,采用人肝微粒体系统的基于探针的测定方法,研究白术内酯Ⅱ对CYP450酶系抑制机理、动力学和抑制类型。结果显示白术内酯Ⅱ代谢受CYP1A2、CYP2C9和CYP3A4抑制剂的影响,最高抑制率分别为41.35%、41.97%和82.45%;白术内酯Ⅱ对P450五种亚型CYP2C9、CYP1A2、CYP2C19、CYP3A4和CYP2D6的IC50值分别为69.7、84.3、92.4、173.8和190.1μmol/L,Ki值分别为190.6、179.1、>200、72.2和66.8;白术内酯Ⅱ对CYP1A2为非竞争性抑制,对CYP2C9和3A4为竞争性抑制,对CYP2C19和2D6为混合型抑制。研究结果表明,CYP1A2、CYP2C9和CYP3A4是参与白术内酯Ⅱ代谢的CYP亚型,白术内酯Ⅱ对5种CYP450亚型具有不同的抑制机制和抑制强度。
基金
National Natural Science Foundation of China(Grant No.81660757)
Jiangxi Provincial Academic
Technical Leader Training Program for Major Disciplines(Grant No.20162BCB22015)
The Science Foundation of Health and Family Planning Commission of Jiangxi Province(Grant No.20181140)。
