多反应监测方法评估丹参和人参对大鼠肝药酶的调控作用  

Evaluation of regulation of Salvia miltiorrhiza and Panax ginseng on rat liver CYP450 using multiple reaction monitoring

作　　者：程汉兴 孙爱华[2] 王宇光[3] 高月[3] 姜颖[1,2] 

机构地区：[1]安徽医科大学,合肥安徽230032 [2]军事医学科学院放射与辐射医学研究所北京蛋白质组研究中心蛋白质组学国家重点实验室,北京102206 [3]军事医学科学院放射与辐射医学研究所药理毒理研究室,北京100850

出　　处：《中草药》2016年第20期3647-3655,共9页Chinese Traditional and Herbal Drugs

基　　金：国家重点基础研究发展计划("973"计划)(2014CBA02001;2011CB505304)

摘　　要：目的建立新型特异性强的高通量细胞色素P450(CYP)丰度测定方法,以人参和丹参为工具药考察药物对肝药酶的调控作用。方法采用基于质谱多反应监测(MRM)模式方法,以串联配体法(QconCAT)表达合成的重标肽段作为内标对丹参与人参处理后,对9种大鼠肝脏CYP(CYP1A1、CYP1A2、CYP2B1、CYP2B2、CYP2C6、CYP2C11、CYP3A1、CYP3A2、CYP17A1)蛋白特异肽段进行定量,探讨丹参和人参对肝脏CYP酶的表达上调或者下调作用。结果 QconCAT重标合成内标肽段与肝脏样品肽段质谱表征一致,与对照组相比,丹参与人参作用后,对大鼠肝内CYP1A1、CYP2B2的表达均具有下调作用,对CYP1A2、CYP2B1的表达均具有上调作用。丹参同时下调CYP3A2、CYP2C11和CYP17A1的表达,而人参则上调这3种蛋白的表达。建立了MRM方法对大鼠肝脏CYP蛋白定量方法,定量方法相对标准偏差在5.9%以内,相对误差在6.8%以内,定量标准曲线的线性系数大于0.9。结论对9种大鼠肝脏CYP酶进行定量研究,其中CYP2B1、CYP2B2和CYP17A1是首次进行蛋白质定量研究。丹参与人参对CYP亚型产生的调控作用存在差异性,为药物配伍实践中提供临床参考价值,避免不良药物相互作用反应。Abstract： Objective To establish a new specific and high-throughput method for CYP450 quantification, and to investigate the effect of drugs on rat liver CYP450 using Salvia miltiorrhiza and Panax ginseng as tool drugs. Methods Mass spectrometry with multiple reaction monitoring （MRM） was applied to quantify nine rat liver enzyme CYP450 isoforms from the rat model with S. miltiorrhiza and P. ginseng. With QconCAT heavy peptides as internal standard, we aimed to assess the effects ofS. miltiorrhiza and P. ginseng on CYP450 isoforms. Results The relative standard deviation and relative error of this method were less than 5.9% and 6.8%, respectively, with good linearity （r2〉0.9）. Nine CYP450s isoforms （CYP1A1, CYP1A2, CYP2B1, CYP2B2, CYP2C6, CYP2Cll, CYP3A1, CYP3A2, and CYP17A1） in rat liver microsome treated with S. miltiorrhiza and P. ginseng were also quantified. Compared with the control, S. miltiorrhiza downregulated the expression levels of CYP 1A1, CYP2B2, CYP3A2, CYP2C 11, and CYP 17A1, while upregulated CYP 1 A2 and CYP2B1. For the effects of P. ginseng, the expression of CYP1A1 and CYP2B2 was decreased, while CYP1A2, CYP2B1, CYP3A2, CYP2Cll, and CYP17A1 were increased. Conclusion We successfully construct a method with MRM to quantify rat liver CYP450 isoforms. And we firstly quantify CYP2B1, CYP2B2, and CYP17A1 in rat liver. The results reveal the regulation of CYP450 by S. miltiorrhiza and P. ginseng, which could provide clinical application for drug compatibility in practice, and avoid adverse drug reactions.

关 键 词：人参 丹参 CYP450 多反应监测 药物配伍 

分 类 号：R285.5[医药卫生—中药学]