六神丸对CYP活性的诱导作用研究  

Study on the Induction Effects of Liushen Pill on the Activity of CYP

作　　者：姜波[1] 张凤[1] 王长虹[2] 陈万生[1] 

机构地区：[1]上海长征医院药材科,上海200003 [2]上海中医药大学中药研究所,上海20120

出　　处：《中国药房》2015年第16期2168-2171,共4页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81325024)

摘　　要：目的:研究六神丸对CYP活性的诱导作用。方法:体外传代培养人肝癌Hep G2细胞。以3、1、0.5μg/ml(以丸质量计,下同)六神丸培养细胞48 h后,采用实时荧光聚合酶链反应(RT-PCR)法测定细胞CYP1A2、CYP2C9、CYP2C19、CYP2D6、CYP2E1、CYP3A4 m RNA的表达;采用Western blot法测定细胞CYP3A4、CYP1A2、CYP2C19、CYP2C9蛋白的表达,采用高效液相质谱法测定咪达唑仑代谢产物1-羟基咪达唑仑的含量,以咪达唑仑生成速率代表CYP3A4活性。上述试验均设阴性对照(0.3%二甲基亚砜)组、阳性对照(30μmol/L利福平、100μmol/L地塞米松)组。将18只Wistar大鼠随机均分为空白对照[等容羧甲基纤维素钠(CMC-Na)]组、六神丸(1.613 mg/kg)组、地塞米松(50 mg/kg)组,连续6 d ig给药后测定大鼠肝微粒1-羟基咪达唑仑生成速率以判定CYP3A1、CYP3A2活性;采用RT-PCR法测定CYP3A m RNA的表达。结果:3、1、0.5μg/ml六神丸培养细胞48 h后,与阴性对照比较,Hep G2细胞中CYP2C9、CYP2C19、CYP2E1、CYP3A4 m RNA表达增强,CYP3A4蛋白表达增强,CYP3A4活性增强,差异均有统计学意义(P<0.05)。与空白对照组比较,大鼠肝微粒体CYP3A1、CYP3A2活性增强,CYP3A1、CYP3A2 m RNA表达增强,差异均有统计学意义(P<0.05)。结论:六神丸对CYP具有一定的诱导作用,其机制可能与增强CYP m RNA的表达作用有关。OBJECTIVE： To study the induction effects of Liushen pill on the activity of CYP. METHODS： Human hepatoma hepG2 cells were subcultured in vitro. After the cells were cultured in 3μg/ml, 1 pg/ml and 0.5 μg/ml Liushen pill solutions （take pill quality plan, the same below） for 48 h, real-time quantitative fluorescent polymerase chain reaction （RT-PCR） technology was used to determine the expression of CYP1A2, CYP2Cg, CYP2C19, CYP2D6, CYP2E1 and CYP3A4 mRNA in cells; Western blot method was respectively employed to determine the expression of CYP3A4 , CYPIA2, CYP2C19, CYP2C9 protein in ceils, and HPLC-MS was adopted to determine the content of metabolite 1-hydroxy midazolam of midazolam, and the activity of CYP3A4 was showed by formation rate of midazolam. There set a negative control group （isometric dimethyl sulfoxide） and posi- tive control groups （30 μmol/L rifampicin and 100 μmol/L dexamethasone）. 18 Wistar rats were randomly divided into blank con- trol group [isometric sodium carboxymethyl cellulose solution （CMC-Na）], Liushen pill group （1.613 mg/kg） and dexamethasone group （50 mg/kg）. After the drugs were administered to these rats for 6 days, ig, the production rates of 1-hydroxymidazolam in rats＇ liver microsomes were determined to judge the activity level of CYP3A; RT-PCR method was adopted to determine the expres- sion of CYP3A1, CYP3A2 mRNA. RESULTS： After the cells were cultured in 3 μg/ml, 1μg/ml and 0.5μg/ml Liushen pill solu- tions for 48 h, compared with negative control group, the expression level of CYP2C9, CYP2C19, CYP2E1 and CYP3A4 mRNA were increased, CYP3A4 protein was increased and the activity of CYP3A4 was increased, with significant difference （P〈0.05）. Compared with blank control group, the activity of CYP3A1, CYP3A2 in rats＇ liver microsomes was increased and the experssion levels of CYP3A1, CYP3A2 mRNA were increased, with significant difference （P〈0.05）. CONCLUSIONS： Liushen pill has cer- tain inductive effects on CYP by a me

关 键 词：六神丸 诱导 人肝癌HEPG2细胞 细胞色素P450 大鼠 

分 类 号：R969.2[医药卫生—药理学]