氯吡格雷对人参皂苷Rg1血浆蛋白结合率的影响

Effect of Clopidogrel on Plasma Protein Binding Rates of Ginsenoside Rg1

目的研究氯吡格雷对人参皂苷Rg1血浆蛋白结合率影响。方法建立人参皂苷Rg1在胎牛血清及透析外液中含量测定方法,随机分成人参皂苷Rg1低、中、高三种浓度单独组(浓度分别为0.4、1.0、5.0 mg/L)和人参皂苷Rg1低、中、高三种浓度合并氯吡格雷组(人参皂苷Rg1浓度依次为0.4、1.0、5.0 mg/L,合并使用2.0 mg/L氯吡格雷),采用平衡透析法观察合并使用氯吡格雷时对人参皂苷Rg1的血清蛋白结合率影响,采用同源模建方法(homology modeling)构建牛血清白蛋白(bovine serum albumin,BSA)三维构建结构,在此基础上采用分子对接方法观察氯吡格雷和人参皂苷Rg1与BSA间结合效应,观察在血浆蛋白结合率层面的相互作用。结果人参皂苷Rg1低、中、高三种浓度单独组与胎牛血清蛋白结合率分别为(11.2±2.1)%、(13.4±2.2)%、(14.6±1.4)%,人参皂苷Rg1在低、中、高三种浓度合并氯吡格雷组血浆蛋白结合率分别下降,分别为(6.5±2.3)%、(9.2±1.5)%、(12.1±1.7)%,差异有统计学意义(P<0.05),分子对接结果显示两个化合物与BSA存在竞争结合效应。结论平衡透析法和分子对接实验综合表明氯吡格雷对人参皂苷Rg1血浆蛋白结合率存在影响作用。

Objective To observe the effect of clopidogrel on plasma protein binding rates of gin- senoside Rgl. Methods Concentrations of ginsenoside Rgl were measured in fetal bovine serum and phosphate buffered solution （PBS）. Samples were randomly divided into ginsenoside Rgl groups （ low： 0.4; middle. 1.0; high 5.0 mg/L, respectively） and clopidogrel combined ginsenoside Rgl groups （low. 0.4 mg/L ＋2.0 mg/L clopidogrel; middle： 1.0 mg/L ＋2.0 mg/L clopidogrel; high.. 5.0 mg/L ＋2.0 mg/L clo- pidogrel）. The effect of clopidogrel on plasma protein binding rates of ginsenoside Rgl was observed u- sing equilibrium dialysis. Then 3-dimensional structure of bovine serum albumin （BSA） was constructed using homology modeling. On this basis, binding effect of small compounds （ginsenoside Rgl and clopi- dogrel~） and BSA was observed using molecular docking method. Results The serum protein binding rate was 11.2% ±2.1% in the low dose ginsenoside Rgl group, 13.4% ±2.2% in the middle dose gin- senoside Rgl group, and 14.6% ±1.4% in the high dose ginsenoside Rgl group, respectively. It was 6.5% ±2.3% in the clopidogrel combined low dose ginsenoside Rgl group, 9.2% ± 1.5% in the clopi- dogrel combined middle dose ginsenoside Rgl group, 12.1% ±1.7% in the clopidogrel combined highdose ginsenoside Rgl group, respectively. They were lower in clopidogrel combined ginsenoside Rgl groups than in ginsenoside Rgl groups with statistical difference （P 〈0.05）. Results of molecular doc- king showed that competitive binding effect existed between compounds （ginsenoside Rgl and clopi- dogrel） and BSA. Conclusion Results of equilibrium dialysis and molecular docking comprehensively in- dicated clopidogrel had effect on plasma protein binding rate of ginsenoside Rgl.