大黄?虫丸中基于血小板P2Y1及P2Y12受体作用的抗血小板聚集活性成分筛选＊

目的 结合分子对接和血小板聚集实验对大黄?虫丸中基于血小板P2Y1及P2Y12受体作用的抗血小板聚集活性成分进行初步筛选。方法 以P2Y1及P2Y12受体作为靶受体,分别与大黄?虫丸中9种主要成分（大黄酚、大黄素、大黄酸、大黄素甲醚、芦荟大黄素、黄芩苷、苦杏仁苷、芍药苷和甘草酸）进行分子对接,计算结合能并分析分子间作用力,与P2Y1及P2Y12受体自带活性配体进行比较。随后以腺苷-5''-二磷酸钠（ADP）为诱导剂,观察大黄?虫丸中9种主要成分在同一浓度下对血小板聚集的作用,以此初步验证分子对接的结果。结果 分子对接结果显示,甘草酸、黄芩苷和大黄酸与P2Y1受体结合能较低,结合位置与自带配体较相似;大黄酸、大黄酚、大黄素、大黄素甲醚和芦荟大黄素与P2Y12受体结合能相对较低,结合位置与自带配体相对较相似。ADP诱导的血小板聚集实验中,2 nmol·L^-1的大黄酸和芦荟大黄素与空白组相比具有显著的抑制作用,聚集率分别为41.3%和73.1%,抑制率为51.2%和13.5%。结论 结合分子对接和血小板聚集实验结果,推测大黄酸和芦荟大黄素可能是大黄?虫丸中基于P2Y1及P2Y12受体通路的抑制血小板聚集的主要活性成分。

Screening of Anti-platelet Aggregation Active Components Based on Platelet P2Y1 and P2Y12 Receptors in Da Huang Zhe Chong Pill

Objective Screen the anti-platelet aggregation active components based on platelet P2Y1 and P2Y12 receptor in Da Huang Zhe Chong Pill by combining molecular docking and platelet aggregation experiment.Methods P2Y1 and P2Y12 receptors were used as target receptors for molecular docking with 9 main components (Chrysophanol, Emodin, Rhein, Physcion, Aloe emodin, Baicalin, Amygdalin, Paeoniflorin and Glycyrrhizic acid) of Da Huang Zhe Chong Pill, respectively. The binding energy was calculated and the intermolecular interaction was analyzed, and compared with the active ligands of P2Y1 and P2Y12 receptors. Then, ADP was used as inducer to observe the effects of 9 main components in Da Huang Zhe Chong Pill on platelet aggregation at the same concentration, so as to preliminarily verify the results of molecular docking.Results The molecular docking results showed that Glycyrrhizic acid, Baicalin And Rhein had low binding energy with P2Y1 receptor, and the binding position was similar to the self-contained ligand. The binding energy of Rhein, Chrysophanol, Emodin, Physcion and Aloe emodin with P2Y12 receptor was relatively low, and the binding position was relatively similar to the self-contained ligand. In ADP induced platelet aggregation experiment, 2 nmol·L^-1 Glycyrrhizic acid, Rhein and Aloe emodin had significant inhibitory effects compared with the blank group. The aggregation precentage were 41.3%, 45.2% and 73.1%, and the inhibition precentage were 51.2%, 46.6% and 13.5%, respectively.Conclusion Combined with the results of molecular docking and platelet aggregation experiment, it is speculated that Glycyrrhizic acid, Rhein and Aloe emodin may be main the active components of Da Huang Zhe Chong Pill to inhibit platelet aggregation based on P2Y1 and P2Y12 receptor pathway.