摘要
目的:采用Cocktail探针药物法研究木犀草素体外对大鼠细胞色素P450(cytochrome P450,CYP450)亚型的影响。方法:建立并优化含有大鼠肝微粒体、探针药物、木犀草素或阳性抑制剂的体外孵育体系,分为空白对照组、木犀草素组、阳性对照组,孵育终止后使用HPLC方法检测各组别代谢产物的生成量,采用GraphPad Prism 9.0计算木犀草素和阳性抑制剂的半数抑制浓度值(IC_(50))。结果:孵育体系经优化,确定大鼠肝微粒体浓度为1.5 mg·mL^(-1),孵育时间为30 min,非那西汀、睾酮、甲苯磺丁脲的K_m值分别为85.62,146.90和78.36μmol·L^(-1)。所建立孵育体系的有效性经已知的阳性抑制剂进行了验证。木犀草素对CYP1A2,CYP3A4和CYP2C9的IC_(50)值分别为19.92,4.64和9.29μmol·L^(-1)。结论:本实验成功地建立了木犀草素的体外孵育体系,木犀草素对大鼠肝微粒CYP1A2,CYP3A4和CYP2C9均具有抑制作用。
Objective:To investigate the potential inhibition of luteoline against CYP1 A2,CYP3 A4 and CYP2 C9 activities in rat liver microsomes through cocktail approach in vitro.Methods:Incubation method was developed and validated for in vitro CYP450 inhibition assay,comprising rat liver microsomes,a cocktail of substrates,luteoline or positive inhibitors.The experiment was divided into blank control group,luteolin group and positive control group.After incubation,the formation of metabolites was investigated by HPLC.The IC50 values of luteolin and positive inhibitors were calculated by GraphPad Prism 9.0.Results:The optimal concentration of rat liver microsomes was 1.5 mg·mL^(-1),and incubation time was 30 min.The Km values of phenacetin,testosterone and tolbutamide were 85.62,146.90 and 78.36μmol·L^(-1),respectively.The established incubation system was validated by known positive inhibitors.The IC50 values of luteolin to CYP1 A2,CYP3 A4,and CYP2 C9 were 19.92,4.64 and 9.29μmol·L^(-1),respectively.Conclusion:An improved cocktail approach was established and validated for CYP inhibition assay.Luteolin has inhibitory activities against CYP1 A2,CYP3 A4 and CYP2 C9.
作者
高琳
宋可心
任纪平
衣洁
李倩
唐景玲
GAO Lin;SONG Ke-xin;REN Ji-ping;YI Jie;LI Qian;TANG Jing-ling(Department of Pharmaceutics,College of Pharmacy,College of Pharmacy,Harbin Medical University,Harbin 150000,China;Department of Pharmaceutical Analysis and Analytical Chemistry,College of Pharmacy,Harbin Medical University,Harbin 150000,China)
出处
《中国新药杂志》
CAS
CSCD
北大核心
2022年第6期590-595,共6页
Chinese Journal of New Drugs
基金
国家自然科学基金项目(81970055)
哈尔滨医科大学药学院杰出青年基金项目(2019-JQ-01和2019-JQ-03)。
