Objective: To explore the main factors of platelet spreading and provide the foundation for related research.
Methods: Platelets (2×107/ml) were draw from C57BL/6J mouse and kept at 22 ℃ for 1-2 hours. Platelets (2×107/ml) were were allowed to adhere and spread on the fibrinogen-coated slides, after staining F-actin in platelets, the platelets were observed with the confocal microscopy. The effects of different concentrations of fibrinogen (10 μg/ml, 30 μg/ml, 100 μg/ml) and kinds of agonists ［thrombin(0.01,0.05,0.1 U/ml), ADP（5,10,20 μmol/L）, U46619（0.125,0.25,0.5 μmol/L）］ on platelets were analyzed. The platelet spreading was successful if the spreading rate was higher after treated with agonists.
Results: Compared to the group which coated with 10 μg/ml and 100 μg/ml fibrinogen, the platelet density is optimal when coated with 30 μg/ml fibrinogen. In addition, under the stimulation of thrombin, ADP and U46619, the spreading rate of platelets showed a certain concentration-dependent increasing.
Conclusion: The platelet spreading is easily influenced by various factors, the platelet spreading can be induced successfully at 0.1 U/ml thrombin, 20 μmol/L ADP and 0.5 μmol/L U46619 on the slide coated with 30 μg/ml fibrinogen.
题目: 血小板铺展的主要影响因素.
目的: 探索血小板铺展的主要影响因素，为相关疾病研究奠定基础.
方法: 体外分离C57BL/6J小鼠的血小板(2×107/ml)，室温静置1-2 h后，将血小板样本均匀的铺在纤维蛋白原包被的载玻片上，利用鬼笔环肽标记骨架蛋白F-actin，封片后通过荧光共聚焦显微镜观察血小板铺展状态，分析不同浓度的纤维蛋白原和激动剂等因素对血小板铺展的影响。实验中纤维蛋白原浓度为10、30和100 μg/ml， 激动剂为凝血酶（0.01、0.05和0.1 U/ml）、ADP（5、10和20 μmol/L）和U46619（0.125、0.25和0.5 μmol/L）。若激动剂处理后的血小板呈树突状并且铺展率较高，则说明血小板铺展的方法构建成功.
结果: 与10和100 μg/ml的纤维蛋白原组相比，在包被了30 μg/ml纤维蛋白原的载玻片上粘附的血小板数目最优；另外，在凝血酶、ADP和U46619的刺激下，血小板铺展与刺激剂浓度呈正相关(ADP: r=0.925， U46619: r=0.937).
结论: 血小板的铺展受到很多因素的影响，30 μg/ml的纤维蛋白原、0.1 U/ml的凝血酶、20 μmol/L的ADP和0.5 μmol/L的U46619可成功诱导2×107/ml血小板的铺展.
Keywords: adhesion; fibrinogen; integrin; spreading.