免疫性血小板减少症气不摄血证小鼠外周血小板活化功能的研究  

Activation of peripheral blood platelets in a mouse model of immune thrombocytopenia with qi failing to control blood syndrome

作　　者：王攀 蔺亚东 武曲星 侯敏 高文静 刘建勋[1] 任钧国[1] WANG Pan;LIN Yadong;WU Quxing;HOU Min;GAO Wenjing;LIU Jianxun;REN Junguo(Beijing Key Laboratory of Pharmacology of Chinese Materia Medica,Institute of Basic Medical Sciences of Xiyuan Hospital China Academy of Chinese Medical Sciences,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所中药药理北京市重点实验室,北京100091

出　　处：《中国比较医学杂志》2020年第7期45-50,共6页Chinese Journal of Comparative Medicine

基　　金：国家重点基础研究发展计划(973)项目(2015CB554405);国家重大新药创制专项(2017ZX09301018)。

摘　　要：目的观察免疫性血小板减少症(immune thrombocytopenia,ITP)气不摄血证小鼠外周血小板活化功能的变化。方法BALB/c小鼠随机分为正常组和模型组,模型组小鼠采用第1周睡眠剥夺,第2、3周睡眠剥夺复合腹腔注射抗血小板血清(anti-platelet serum,APS)的方法复制ITP气不摄血证小鼠模型。实验结束后,扫描电镜观察小鼠血小板形态,ELISA检测血小板α颗粒(β-TG、PF-4)、致密颗粒(ATP、ADP)的释放情况,流式细胞术检测小鼠血小板CD62P的表达率、血小板-中性粒细胞黏附(PNA)的百分率、血小板线粒体膜电位(MMP)和活性氧平均荧光强度(ROS-MFI)。结果正常组小鼠血小板大多呈圆形或圆盘形,有少量伪足形成,模型组小鼠血小板大多形状不规则,有大量伪足形成,部分黏附聚集成团。与正常组相比,模型组小鼠血小板β-TG、PF-4、ATP、MMP、ROS-MF明显升高(P<0.01),CD62P、ADP、PNA明显升高(P<0.05)。结论ITP气不摄血证小鼠外周血小板呈高度活化状态且凋亡增加。Objective To observe the activation function of peripheral blood platelets in a mouse model of immune thrombocytopenia(ITP)with qi failing to control blood syndrome.Methods BALB/c mice were randomly divided into control and model groups.The model group was subjected to sleep deprivation for one week,followed by two weeks of sleep deprivation combined with intraperitoneal injection of anti-platelet serum to replicate an ITP mouse model with qi failing to control blood syndrome.At the end of the experimental period,the morphology of platelets isolated from the mice was observed by a scanning electron microscope.Moreover,ELISA was used to detect the release of platelet alpha particles(β-TG,PF-4)and dense granules(ATP,ADP).Flow cytometry was used to detect CD62P expression,plateletneutrophil aggregation(PNA),mitochondrial membrane potential(MMP),and the mean fluorescence intensity of reactive oxygen species(ROS-MFI).Results The platelets from the control group were mostly round or disk-like with a small degree of pseudopodia formation,while platelets from the model group were mostly irregular in shapes,with a large degree of pseudopodia formation,and some of them adhered and aggregated into groups.Compared with the control group,β-TG,PF-4,ATP,MMP,and ROS-MFI in the model group were significantly increased(P<0.01),while CD62P,ADP,and PNA were increased significantly,but to a lesser extent(P<0.05).Conclusions Platelets from the peripheral blood of a mouse model of ITP with qi failing to control blood syndrome were highly activated and exhibited increased rates of apoptosis compared with control mice.

关 键 词：ITP气不摄血证 血小板 活化功能 

分 类 号：R-33[医药卫生]