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

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

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。

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 RSA were（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.