齐澳 任灿 李俊 赵筱萍 金强 何江敏

摘要 目的：使用轉基因斑马鱼模型筛选冠心宁片中具有抗血栓活性的组分及药效物质，并探讨其作用机制。方法：将斑马鱼胚胎分为对照组、造模组、给药组及阳性药组，使用苯肼诱导血栓模型。通过荧光显微成像结合血红蛋白染色对冠心宁片各组分抗血栓作用进行定性和定量分析。进一步探寻活性组分中起抗血栓作用的主要成分。最后进行活性氧（ROS）检测及脂质氧化损伤丙二醛（Malondialdehyde，MDA）分析，探讨其抗血栓机制。结果：实验结果显示冠心宁片水溶性组分可显著减少外周血红细胞聚集（P<0.01），增加回心血量，且效果呈剂量依赖性。因多糖为冠心宁片水溶性成分中的主要物质，进一步实验发现丹参多糖和川芎多糖也具有抗血栓活性，且能显著降低斑马鱼的氧化应激水平，丹参多糖还具有下调胚胎脂质过氧化水平的作用。结论：丹参多糖和川芎多糖为冠心宁片水溶性组分中具有抗血栓作用的活性化合物，可能与其抑制氧化应激作用有关，具有开发为抗血栓治疗先导物的潜在应用前景。

关键词 血栓;斑马鱼;丹参多糖;川芎多糖;氧化应激;苯肼

Study on Antithrombotic Activity of Water-soluble Components in Guanxinning Tablet

QI Ao1，REN Can1，LI Jun1，ZHAO Xiaoping2，JIN Qiang3，HE Jiangmin3

（1 College of Pharmaceutical Sciences，Zhejiang University，Hangzhou 310058，China; 2 School of Basic Medical Sciences，Zhejiang Chinese Medical University，Hangzhou 310053，China; 3 Chiatai Qingchunbao Pharmaceutical Co.，Ltd.，Hangzhou 310023，China）

Abstract Objective：To screen the fractions and pharmacodynamic substances with antithrombotic activity in Guanxinning Tablet by using transgenic zebrafish model，and to explore the mechanism of action.Methods：Zebrafish embryos were divided into a control group，a model group，an administration group and a positive group，and the thrombus model was induced by phenylhydrazine.Qualitative and quantitative analysis of the antithrombotic effect of each component of Guanxinning tablets was carried out by fluorescence microscopy imaging combined with hemoglobin staining.Further explore the main components of the active ingredients that play an anti-thrombotic effect.Finally，ROS measurement and MDA analysis were performed to explore its anti-thrombotic mechanism.Results：The experimental results showed that the water-soluble components of Guanxinning Tablets can significantly reduce peripheral red blood cell aggregation （P<0.01），increase the return blood volume，and the effect was dose-dependent.Because polysaccharides were the main substances in the water-soluble components of Guanxinning Tablets，further experiments have found that Danshen polysaccharides and Ligusticum chuanxiong polysaccharides also had antithrombotic activity and could significantly reduce the oxidative stress level of zebrafish.Conclusion：Danshen polysaccharides and Ligusticum chuanxiong polysaccharides are active compounds with antithrombotic effects in the water-soluble components of Guanxinning tablets，which may be related to their inhibitory effects on oxidative stress and have potential application prospects for the development of antithrombotic therapies.

Keywords Blood clots; Zebrafish; Salvia miltiorrhiza polysaccharide; Ligusticum chuanxiong polysaccharide; Oxidative stress; Phenylhydrazine

中图分类号：R285.5文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.23.006

近30年来，心血管疾病仍居全球死亡因素的首位[1]，超過了总死亡原因构成的40%，远高于肿瘤及其他疾病的死亡率[2]。血栓形成是冠心病等主要心血管疾病的关键病理过程。按照形成部位，血栓可分为动脉血栓和静脉血栓。动脉粥样硬化斑块的破裂可导致动脉阻塞，从而引发心脏病发作、中风或猝死等一系列疾病。……