基于液相色谱质谱联用氯吡格雷-人参皂苷血浆蛋白结合率协同效应

目的研究大鼠血浆中氯吡格雷-人参皂苷血浆蛋白结合率协同效应。方法选择人参皂苷Rg1、Rb1为指标，首先建立了
Rg1、Rb1在透析内液（大鼠血清RSA）、外液（PBS）含量检定方法，采用平衡透析法揭示了氯吡格雷对人参Rg1、Rb1血清蛋白结
合率协同作用；接下来，采用同源模建方法建构RSA三维构造，并用分子模拟方法揭示氯吡格雷、人参皂苷Rg1、Rb1与大鼠血
清白蛋白RSA间结合效应，揭示氯吡格雷-人参皂苷在血浆蛋白结合率层面协同效应。结果人参皂苷Rg1、Rb1在0.40、1.00、
2.00 mg·L-1三种浓度下与大鼠血清蛋白结合率分别为30.16±2.82、33.42±4.21、34.61±3.42 和50.13±2.34、51.23±3.23、53.11±
3.26，当合并加入氯吡格雷（1.0 mg·L-1）后血浆蛋白结合率分别下降至22.13±2.72、21.42±3.22、25.45±3.52和40.13±3.24、41.25±
4.15、43.11±3.31，分子模拟结果显示氯吡格雷、Rg1、Rb1与RSA存在不同程度的竞争结合效应。结论平衡透析法和分子模拟
实验综合揭示了氯吡格雷-人参皂苷血浆蛋白结合率协同效应。

Effect of clopidogrel on plasma protein binding rate of ginsenosides: a liquidchromatography-mass spectrometry-based study

Objective To investigate the effect of clopidogrel on the binding rate of ginsenosides with rat serum proteins (RSA).
Methods Equilibrium dialysis and liquid chromatography-mass spectrometry were employed to quantify the concentration of
ginsenoside Rg1 and Rb1. The protein-binding rates of Rg1 and Rb1 in the presence or absence of clopidogrel (1.0 mg/L) were
determined. A molecular simulation model (consisting of homology modeling and molecular docking interaction) was used to
reveal the target protein-compound interactions. Results The binding rates of ginsenosides Rg1 (0.4, 1.0, and 2.0 mg/L) with
RSAwere (30.16±2.82)%, (33.42±4.21)%, and (34.61±3.42)%, and those of and Rb1 were (50.13±2.34)%, (51.23±3.23)%, and (53.11±
3.26)%, respectively. In the presence of clopidogrel, the binding rates of Rg1 decreased to (22.13±2.72)%, (21.42±3.22)%, and
(25.45 ± 3.52)% , and those of Rb1 to (40.13 ± 3.24)% , (41.25 ± 4.15)% , and (43.11 ± 3.31)% , receptively. The molecular docking
suggested that these compounds competed to bind with RSA. Conclusion Clopidogrel can competitively bind to RSA with
ginsenosides to lower the plasma protein binding rates of ginsenosides.