混合探针底物法测定五味子成分对大鼠肝脏细胞色素P450同工酶的影响

Effects of Schisandra chinensis(Wuweizi) constituents on the activity of hepatic microsomal CYP450 isozymes in rats detected by using a cocktail probe substrates method

研究大鼠多次口服五味子成分和体外对肝脏细胞色素P450酶(CYP450)6种同工酶的影响。采用超速离心法制备正常及多次口服五味子醇/水提物的大鼠肝微粒体并与探针药进行体外温孵,应用液相色谱?串联质谱分析方法测定CYP450的6种同工酶特异性探针底物非那西丁(CYP1A2)、右美沙芬(CYP2D2)、双氯芬酸钠(CYP2C6)、美芬妥英(CYP2C11)、氯唑沙宗(CYP2E1)、咪达唑仑(CYP3A1/2)在大鼠肝微粒体的代谢产物生成(对乙酰氨基酚、右啡烷、4-羟基双氯芬酸钠、4-羟基美芬妥英、6-羟基氯唑沙宗、1-羟基咪达唑仑)以反映各CYP450同工酶活性。五味子醇提物(28～120μg.mL-1)体外对大鼠肝微粒体CYP450的6个同工酶均有不同程度的抑制作用。大鼠多次口服五味子醇提物(1.5 g.kg-1,qd×7d)对肝脏CYP3A1/2和CYP2E1有显著诱导作用,对CYP2D2和CYP2C11有明显抑制作用,而对CYP2C6和CYP1A2无明显影响。五味子水提物(100～500μg.mL-1)体外对大鼠肝脏CYP450同工酶亦有抑制作用;体内多次给药(1.5 g.kg-1,qd×7d)对肝脏CYP2D2有明显抑制作用,对CYP2E1有显著诱导作用,但对CYP2C6、CYP3A1/2、CYP1A2、CYP2C11无明显影响。提示五味子成分体内外对CYP450同工酶具有不同程度的诱导或抑制作用,是否与合用药物产生相互作用可能主要取决于参与药物体内处置的CYP450亚型及其比例,临床应用时应密切注意。

Effects of constituents from Schisandra chinensis（Wuweizi） on six liver microsomal CYP450 isozymes（CYP1A2,CYP2C6,CYP2C11,CYP2D2,CYP2E1 and CYP3A1/2） were studied in rats in vivo and in vitro.The in vitro incubation was conducted using liver microsomes of rats after multiple dosing of alcoholic/water extract from Schisandra chinensis.A HPLC-MS method was applied to determine the metabolites formation of six CYP450s probe substrates（phenacetin-CYP1A2,dextromethorphan-CYP2D2,diclofenac sodium-CYP2C6,mephenytoin-CYP2C11,chlorzoxazone-CYP2E1 and midazolam-CYP3A1/2） in rat liver microsomal incubations.The activity of CYP450 isozymes were represented by the formation of metabolites.Alcoholic extract of Schisandra chinensis（28-120 μg？mL-1） showed significant inhibitory effect on six CYP450 isozymes to a certain extent in vitro.Multiple dosing of Schisandra chinensis alcoholic extract（1.5 g-kg-1,qd×7d） had significant induction on CYP2E1 and CYP3A1/2,inhibition on CYP2D2 and CYP2C11,and no effect on CYP2C6 and CYP1A2.Water extract of Schisandra chinensis（100-500 μg-mL-1） also exhibited inhibition on the activity of CYP450 isozymes in vitro,whereas multiple administrations（1.5 g-kg-1,qd×7d） had significant induction of CYP2E1 and inhibition on CYP2D2,no effect on CYP2C6,CYP3A1/2,CYP1A2 or CYP2C11.The results suggested that the constituents from Schisandra chinensis exhibited the inhibition and induction on six rat liver microsomal CYP450 isozymes to a certain extent in vivo and in vitro.The possibility of interaction between Schisandra chinensis and coadministrative drugs will be considered base on the levels and subtype of CYP450 involved in the drug metabolism.