基于功效实验-网络药理学-HPLC探讨大黄-桃仁配伍活血化瘀的物质基础及分子作用机制

Study on the material basis and molecular mechanism of Rhei Radix et Rhizoma−−Persicae Semen combination in activating blood circulation and dispelling blood stasis based on efficacy experiments,network pharmacology and HPLC

本研究整合功效实验、网络药理学和HPLC探讨大黄-桃仁药对(Rhei Radix et Rhizoma-Persicae Semen combination,RRR-PS)配伍发挥活血化瘀作用的功效物质群及分子作用机制。建立大鼠血瘀证模型,进行血液流变指标和凝血四项综合评价,结果显示,与模型组相比,RRR-PS组大鼠的全血黏度、血沉、红细胞聚集指数等指标均明显回调(P<0.01)。网络药理学发现RRR-PS通过作用于钙调蛋白1(CALM1)、一氧化氮合酶2(NOS2)、糖皮质激素受体(NR3C1)等靶点,调控血小板活化、过氧化物酶体增殖物激活受体(PPAR)、血管内皮生长因子(VEGF)等信号通路发挥活血化瘀的功效,并发现关键成分:番泻苷B、儿茶素、大黄素、大黄酚、大黄酸、芦荟大黄素、大黄素甲醚、没食子酸。运用HPLC探究关键成分的溶出率,结果发现RRR-PS配伍后,儿茶素、大黄素、大黄酚、大黄素甲醚4种主要功效物质含量均显著提高(P<0.01)。综上所述,RRR-PS活血化瘀功效显著,其作用机制与促进血管生成、抗凝血、抗血栓、抗炎有关;功效发挥与RRR-PS配伍后,溶剂系统改变,儿茶素、大黄素、大黄酚、大黄素甲醚大量溶出相关。研究结果可进一步为后续RRR-PS配伍活血化瘀功效物质及作用机制研究提供参考。动物实验已获得陕西中医药大学实验动物委员会批准(批准号:SUCMDL20210309002)。

In this study,the effective substance group and molecular mechanism of Rhei Radix et Rhizoma-Persicae Semen combination(RRR-PS)in activating blood circulation and dispelling blood stasis were investigated by integrating efficacy experiments,network pharmacology and HPLC.The rat model of blood stasis syndrome was established,and the blood rheology index and coagulation four comprehensive evaluation were carried out.The results showed that compared with the model group,the whole blood viscosity,erythrocyte sedimentation rate and erythrocyte aggregation index of the rats in the RRR-PS group were significantly callback(P<0.01).Network pharmacology found that RRR-PS combination exerted the effect of activating blood circulation and dispelling blood stasis by acting on calmodulin-1(CALM1),nitric oxide synthase-2(NOS2),glucocorticoid receptor(NR3C1)and other targets,regulating platelet activation,peroxisome proliferator-activated receptor(PPAR),vascular endothelial growth factor A(VEGF)and other signaling pathways,and found key components:sennoside B,(+)-catechin,emodin,physcion,rhein,aloe-emodin,chrysophanol,gallic acid.HPLC was used to explore the dissolution rate of key components.The results showed that the contents of catechin,emodin,chrysophanol and physcion were significantly increased after RRR-PS combination(P<0.01).In summary,RRR-PS has a significant effect on promoting blood circulation and removing blood stasis,and its mechanism of action is related to promoting angiogenesis,anti-coagulation,anti-thrombosis and anti-inflammation.The efficacy is related to the change of solvent system and the large dissolution of catechin,emodin,chrysophanol and physcion after the RRR-PS combination.The results of the study can further provide a reference for the follow-up study on the active substances and mechanism of the RRR-PS combination.Animal experiments have been approved by the Experimental Animal Committee of Shaanxi University of Traditional Chinese Medicine(No.SUCMDL20210309002).