斑马鱼血栓模型在中药活性成分筛选中的适用性  

Applicability of a zebrafish thrombosis model in screening active ingredients of traditional Chinese medicine

作　　者：莫彩莲 李杰 王加珍 刘鑫 林圣华 牟艳玲 张云 刘可春 何秋霞 MO Cai-lian;LI Jie;WANG Jia-zhen;LIU Xin;LIN Sheng-hua;MOU Yan-ling;ZHANG Yun;LIU Ke-chun;HE Qiu-xia(Engineering Research Center of Zebrafish Model for Human Diseases and Drug Screening of Shandong Province,Biology Institute, Qilu University of Technology (Shandong Academy of Sciences),Jinan 250014, China;Institute of Materia Medica, Shandong First Medical University(Shandong Academy of Medical Sciences), Jinan 250062, China)

机构地区：[1]齐鲁工业大学(山东省科学院)山东省科学院生物研究所,山东省人类疾病斑马鱼模型与药物筛选工程技术研究中心,山东济南250014 [2]山东第一医科大学(山东省医学科学院)药物研究所,山东济南250062

出　　处：《山东科学》2021年第4期52-59,共8页Shandong Science

基　　金：山东省重大科技创新工程(2019JZZY021020)。

摘　　要：为了考察斑马鱼血栓生成模型在中药成分活性筛选中的适应性,利用体外血小板聚集实验和体内斑马鱼血栓形成实验,分别检测中药单体对血小板聚集和血栓形成的影响。筛选模型选择3~5 d的野生型AB系斑马鱼以及转基因TG系(integrinαIIb:EGFP)血小板标记绿色荧光斑马鱼,利用三氯化铁诱导血栓形成,以凝血时间为指标,用20种中药单体成分处理斑马鱼,考察对斑马鱼血栓形成的影响。同时,采集大鼠全血制备富血小板血浆和贫血小板血浆,利用血小板聚集仪测定血小板聚集率,检测中药单体成分活性。结果显示:有7种中药单体成分具有抑制斑马鱼血栓形成活性,首次发现桃叶珊瑚苷具有明显的抑制斑马鱼血栓形成作用;体外血小板聚集实验结果显示有12种中药材成分具有抑制血小板聚集的作用,其中100μg/mL红景天苷的血小板聚集抑制率88.22%;利用斑马鱼血栓模型检测出的7种中药单体与体外血小板聚集实验结果一致。斑马鱼血栓生成模型对中药抗血栓活性筛选具有良好的适应性。This study is aimed to investigate the applicability of zebrafish thrombosis model in the screening of active ingredients of traditional Chinese medicine.In vitro platelet agglutination experiments and in vivo zebrafish thrombosis experiments were performed to detect the effects of monomers in traditional Chinese medicine material on platelet aggregation and thrombosis.We selected a 3~5-days-old transgenic TG line(integrinαIIb:enhanced green fluorescent protein)of platelet-labeled green fluorescent zebrafish and wild-type AB line zebrafish as screening models and used ferric chloride to induce thrombosis.The zebrafish were treated with 20 types of monomer components of traditional Chinese material,and clotting time was used as an indicator to investigate the effect of these samples on thrombosis in zebrafish.Simultaneously,the whole blood of rats was collected to prepare platelet-rich plasma and platelet-poor plasma,and platelet aggregation rate was measured by a platelet agglutination instrument to detect the activity of monomer components of Chinese medicine material.There were seven types of monomer components of traditional Chinese medicine material demonstrated antithrombotic effects in zebrafish.For the first time,it was first found that aucubin had a significant inhibitory effect on thrombosis in zebrafish.The results of in vitro platelet aggregation test showed that 12 traditional Chinese medicine components inhibited platelet aggregation,of which the maximum inhibition of platelet aggregation(88.22%)was exhibited by 100μg/mL salidroside.The seven types of monomer components of traditional Chinese medicine material found to have antithrombotic effects in zebrafish model exhibited good results in in vitro platelet aggregation experiments as well.The zebrafish thrombosis model has good adaptability to the screening of antithrombotic activity of traditional Chinese medicine.

关 键 词：斑马鱼 血栓形成 中药 药物筛选 先导化合物 

分 类 号：R965.1[医药卫生—药理学]