潘洁,秦兰,杨淑婷,巩仔鹏,李勇军,陆苑
PAN Jie,QIN Lan,YANG Shuting,GONG Zipeng,LI Yongjun,LU Yuan

• 1:贵州医科大学民族药与中药开发应用教育部工程研究中心&省部共建药用植物功效与利用国家重点实验室
• 2:贵州医科大学药学院
• 3:贵州医科大学贵州省药物制剂重点实验室

摘要(Abstract)：

目的:考察羊耳菊(YEJ)提取物对大鼠和人肝微粒体CYP450酶的6种主要亚型酶的抑制作用。方法:采用钙盐沉淀法提取肝微粒体,在肝微粒体孵育体系中,用PBS溶液将YEJ提取物逐级稀释至5个不同浓度(0.01、0.05、0.25、0.5、1 g/L),分别与混合探针底物(非那西丁、氯唑沙宗、右美沙芬、奥美拉唑、甲苯磺丁脲、咪达唑仑和睾酮)共同孵育,采用超高效液相色谱串联质谱法(UPLC-MS/MS)同时测定6种探针底物对应代谢物的生成量,以空白溶剂组的代谢物生成量为V_0,其余不同YEJ提取物浓度的代谢物生成量为Vx,求得剩余酶活性I=(1-Vx/V0)×100,通过GraphPad v 5.0软件拟合曲线,并计算IC_(50)值。结果:YEJ提取物对大鼠肝微粒体CYP1A2、CYP2C11、CYP2C19、CYP2D4、CYP2E1和CYP3A2的IC_(50)值范围为0.287～152.407 g/L,对人肝微粒体CYP1A2、CYP2C9、CYP2C19、CYP2D6、CYP2E1和CYP3A4的IC_(50)值范围为0.258～19.275 g/L。结论:YEJ提取物对大鼠和人肝微粒体中6种亚型酶活性存在较弱的抑制作用。
Objective: To investigate the inhibitory effects of Inula cappa(YEJ) extract on the activities of six CYP450 enzyme isoforms in rat and human liver microsomes. Methods: Liver microsomes were extracted by calcium salt precipitation. In the liver microsome incubation system, YEJ extract was diluted to 5 different concentrations(0.01, 0.05, 0.25, 0.5, 1 g/L) with phosphate buffer saline(PBS) buffer, and then incubated with a mixed probe substrates(phenacetin, chlorzoxazone, dextromethorphan, omeprazole, tolbutamide, midazolam, and testosterone). The metabolite amount was determined by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). The amount of metabolites produced by the blank solvent group was recorded as V0, and the amount of metabolites produced by the other different YEJ extract concentrations was recorded as Vx. Residual enzyme activity(I) was calculated using the equation I=(1-Vx/V0)×100, and the activity curve was fitted by GraphPad v 5.0 software. IC_(50) values were calculated based on the activity curve. Results: The IC_(50) values of YEJ extracts ranged from 0.287 to 152.407 g/L on rat liver microsomeal CYP1 A2, CYP2 C11, CYP2 C19, CYP2 D4, CYP2 E1 and CYP3 A2, and the IC_(50) values range from 0.258 to 19.275 g/L on human liver microsomes. Conclusion: YEJ extract has a weak inhibitory effect on the activity of six isoforms of CYP450 in rat and human liver microsomes.

关键词(KeyWords)： 细胞色素P450酶;抑制作用;药物相互作用;人肝微粒体;大鼠肝微粒体;羊耳菊提取物
cytochrome P450 enzyme;inhibition;drug interaction;human liver microsomes;rat liver microsomes;Inula cappa extract

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81860734;U1812403);; 中央引导地方科技专项项目[黔科中引地(2018)4006)];; 贵州省科技厅人才团队项目[黔科合平台人才(2016)5613\5677]

作者(Author): 潘洁,秦兰,杨淑婷,巩仔鹏,李勇军,陆苑
PAN Jie,QIN Lan,YANG Shuting,GONG Zipeng,LI Yongjun,LU Yuan

DOI: 10.19367/j.cnki.1000-2707.2019.11.006

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