附子人参配伍对CYP1A2和CYP3A4酶活性的影响
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摘要
目的:通过建立"Cocktail"探针药物法及体外肝微粒体孵育体系,测定大鼠肝微粒体中CYP1A2和CYP3A4酶的活性,探索附子配伍人参对附子主要成分乌头类生物碱的代谢酶CYP1A2和CYP3A4酶活性的影响。方法:采用"Cocktail"探针药物法,利用CYP1A2和CYP3A4酶的特异性探针底物咖啡因和咪达唑仑,建立同时测定两种探针底物的体外肝微粒体孵育体系;采用RP-HPLC法测定两种探针底物在大鼠肝微粒体孵育体系中的浓度,计算CYP1A2和CYP3A4酶活性的变化。结果:与空白组比较,附子组、人参组、附子人参组对CYP1A2酶活性没有影响,附子组、附子人参组对CYP3A4酶活性没有影响,人参组对CYP3A4酶活性有抑制作用,进而减慢附子主要药效成分乌头类生物碱的代谢,因此,从代谢酶角度可以阐释,附子人参配伍能够增强附子的药效。
The activity of CYP1A2 and CYP3A4 enzymes in rat liver microsomes was determined by establishing "Cocktail" probe drug method and liver microsome incubation system in vitro, and the effects of compatibility of Fuzi and Renshen on the activities of CYP1A2 and CYP3A4 enzymes, which are the main components of Aconitum alkaloids. Methods: Using "Cocktail" probe drug method, and using specific probe substrates of CYP1A2 and CYP3A4 enzymes, caffeine and midazolam, a system for simultaneous determination of two probe substrates in vitro liver microsome incubation system was established. RP-HPLC method was used to determine the concentration of two probe substrates in rat liver microsome incubation system, and the changes of CYP1A2 and CYP3A4 enzyme activities were calculated. Results: Compared with the blank group, the activity of CYP1A2 was not affected in the Fuzi group, Rensen group and compatibility group, but not in the Fuzi group and compatibility group. The Renshen group inhibited the activity of CYP3A4 and slowed down the metabolism of aconite alkaloids, which is the main active ingredient of aconite. Therefore, from the point of view of metabolic enzymes, it can be explained that the compatibility of Fuzi and Renshen can enhance the activity Effectiveness of Fuzi.
The activity of CYP1A2 and CYP3A4 enzymes in rat liver microsomes was determined by establishing "Cocktail" probe drug method and liver microsome incubation system in vitro, and the effects of compatibility of Fuzi and Renshen on the activities of CYP1A2 and CYP3A4 enzymes, which are the main components of Aconitum alkaloids. Methods: Using "Cocktail" probe drug method, and using specific probe substrates of CYP1A2 and CYP3A4 enzymes, caffeine and midazolam, a system for simultaneous determination of two probe substrates in vitro liver microsome incubation system was established. RP-HPLC method was used to determine the concentration of two probe substrates in rat liver microsome incubation system, and the changes of CYP1A2 and CYP3A4 enzyme activities were calculated. Results: Compared with the blank group, the activity of CYP1A2 was not affected in the Fuzi group, Rensen group and compatibility group, but not in the Fuzi group and compatibility group. The Renshen group inhibited the activity of CYP3A4 and slowed down the metabolism of aconite alkaloids, which is the main active ingredient of aconite. Therefore, from the point of view of metabolic enzymes, it can be explained that the compatibility of Fuzi and Renshen can enhance the activity Effectiveness of Fuzi.
引文
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[6]熊亮,彭成,繆璐琳,等.基于“毒-效整合分析思路”探讨有毒中药附子的物质基础[J].世界中医药,2017,12(11):2568-2578.
[2]贺抒,戴鸥,刘建林,等.附子水溶性生物碱治疗急性心力衰竭的研究[J].中药药理与临床,2014,(2):89-92.
[3]Janne T Backman,Marika T Schroder,Pertti J Neuvonen.Effects of gender and moderate smoking on the pharmacokinetics and effects of the CYP1A2 substrate tizanidine[J].European Journal of Clinical Pharmacology,2008,64(1):17-24.
[4]WANG Y,WANG S,LIU Y,et al.Characterization of metabolites and cytochrome P450 isoforms involved in the microsomal metabolism of aconitine[J].J Chromatogr B,2006,844(2):292.
[5]张斐斐,魏飞跃.浅析历代医家对附子配伍的临床应用[J].河南中医,2017,37(9):1662-1666.
[6]熊亮,彭成,繆璐琳,等.基于“毒-效整合分析思路”探讨有毒中药附子的物质基础[J].世界中医药,2017,12(11):2568-2578.