益气活血中药配伍与双抗血小板药物相互作用的机理研究

Supplementing qi and activating blood circulation (SQAB) is a commonly used method for the treatment of cardiovascular and cerebrovascular thrombotic diseases in clinical practice of traditional Chinese medicine (TCM). Anti-thrombosis without increasing the risk of bleeding is the purpose of the treatment of these diseases. However, there are no evidence on whether TCM for SQAB combining with dual-antiplatelet drugs (aspirin, clopidogrel) could further improve the antithrombotic effect and reduce the risk of bleeding. In the present study, the experimental rat model with carotid artery thrombosis (focal cerebral ischemia model) or myocardial infarction, and endothelial injury model induced by lipid oxidation and platelet activation will be established to test the advantage of combining treatment on anti-thrombosis without increasing of bleeding, and to explore the multi-directional molecular mechanism through cyclooxygenase- prostacyclin and ADP-PI3K/Akt, which were well known as the key pathway of the thrombosis or bleeding. Therefore, this study will provide a experimental foundation to support the clinical application of TCM for SQAB combining with dual-antiplatelet drugs, which will be significant to improve the secondary prevention of cardiovascular and cerebrovascular diseases with TCM.

益气活血是中医治疗心脑血管血栓性疾病的常用治法，抗血栓形成同时减少出血风险是心脑血管血栓性疾病治疗的主要目的。益气活血中药配伍联合双抗血小板药物（阿司匹林、氯吡格雷），是否可进一步加强抗血栓形成作用，同时减少出血风险，目前尚缺少研究报道。针对这一问题，本项目通过大鼠颈动脉血栓模型（局部脑缺血模型）、大鼠心肌梗死模型、脂质过氧化和血小板活化致内皮损伤模型，观察益气活血配伍联合双抗抗血栓形成和减少出血风险的优势，并从环氧化酶-血栓素/前列环素和ADP-PI3K/Akt两个抗血小板关键通路，研究益气活血配伍合用双抗发挥双向作用的细胞分子机理，以为益气活血配伍联合双抗干预心脑血管血栓性疾病的临床应用提供实验依据。这对提高中医药心脑血管疾病二级预防疗效，具有重大意义。

本项目通过建立大鼠颈动脉血栓、急性心肌梗死、脑缺血再灌注（I/R）模型及脂质过氧化血小板活化致内皮损伤模型，证实了益气（西洋参茎叶总皂苷，PQS）活血（三七总皂苷，PNS）中药配伍联合双抗抗血栓形成、改善微循环、保护胃黏膜、减少出血风险的优势并揭示了部分机理。. 在体实验证实，益气活血中药联合双抗较单用双抗治疗：①进一步减轻血栓重量，机制与抑制血小板聚集、调节纤溶系统平衡有关；②进一步降低大鼠心肌梗死面积，并增加抗血栓作用，机制与增强阿司匹林（ASA）对血小板环氧化酶（COX）-1/血栓素（TXA2）的抑制，改善TXA2/ 前列环素（PGI2）平衡、激活纤溶系统有关；③进一步改善脑I/R后脑微循环障碍，抑制脑组织坏死，其机制与激活磷脂酰肌醇-3激酶（PI3K）/蛋白激酶B（Akt）Akt/ Ras相关的C3肉毒杆茜毒素底物1（Rac1）通路进一步抑制RhoA/ROCK通路表达以及直接抑制RhoA/ROCK通路表达有关；④减轻双抗所致的胃黏膜损伤，其机制可能与升高血清胃泌素（GAS）、胃动素（MTL）含量和胃组织COX-1、COX-2、前列腺素E-2（PGE2）、血管内皮生长因子（VEGF）、６-酮-前列腺素F1α（6-keto-PGF1α）表达，抑制胃组织肿瘤坏死因子-α（TNF-α）表达有关。. 离体实验证实：益气活血配伍双抗可抑制内皮细胞凋亡和内皮损伤诱导的血小板粘附，其机制与促进内皮细胞PI3K/Akt通路、COX-2/PGI2通路和抑制血小板PI3K/Akt通路、COX-1/TXA2通路有关。PNS抑制血小板粘附的作用较ASA更显著，其机制与PNS抑制血小板COX-1/TXA2通路及促进内皮细胞COX-2/PGI2通路有关。. 本项目为益气活血配伍联合双抗干预心脑血管血栓性疾病提供了可靠的实验依据。这对提高中医药心脑血管疾病二级预防疗效，具有重大意义。