UPLC-MS/MS结合多探针底物方法研究刺五加叶中黄酮苷类成分对CYP450活性的影响

利用超高效液相色谱-串联质谱联用(UPLC-MS/MS)的多反应监测(MRM)技术结合多探针底物方法, 研究了刺五加叶中的主要黄酮苷类化合物槲皮苷、金丝桃苷及芦丁对肝细胞色素P450酶(CYP450)亚型CYP1A2, CYP2C, CYP2E1, CYP2D和CYP3A活性的影响. 结果表明, 3种化合物对各CYP亚型酶均有抑制作用, 其中金丝桃苷和槲皮苷对CYP1A2催化的非那西丁的O-脱乙基反应抑制的IC₅₀值分别为46.53和49.75 μmol/L, 金丝桃苷和芦丁对CYP2E1催化的氯唑沙宗的6-羟基化反应抑制的IC₅₀值分别为99.87和86.36 μmol/L. 机理性抑制实验结果表明, 3种化合物对2种亚型酶的抑制作用是随着预孵时间延长而增强的机理性抑制.

Effects of Flavonoides from the Leaves of Acanthopanax on the Activity of CYP450 Isozymes in Rat Liver Microsomes by a UPLC-MS/MS and Cocktail Probe Substrates Method

Effects of flavonoides including quercitrin, hyperoside and rutin from the leaves of Acanthopanax on the activity of hepatic microsomal CYP450 isozymes(CYP1A2, CYP2C, CYP2E1, CYP2D and CYP3A) in rat liver microsomes were analyzed by a ultra performance liquid chromatography-tandem mass spectrometry(multiple reaction monitoring)UPLC-MS/MS(MRM)] combined with cocktail probe substrates method. The results showed that three compounds on the activities of some CYP450 isozymes were the inhibitory effects using rat liver microsomes, in which the IC50 values of quercitrin and hyperoside to the inhibition of rat microsomal CYP1A2 activity were 46.53 and 49.75 μmol/L, respectively, the IC50 values of hyperoside and rutin for the inhibition of CYP2E1 activity were 99.87 and 86.36 μmol/L, respectively. Mechanism of inhibition experimental results show that three compounds on each CYP450 isozymes inhibiting ability was increased with the increasing of the preincubation time, therefore, the inhibitory effects was a mechanism-based inhibition.