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CD226基因敲除小鼠血小板的结构异常与功能障碍 被引量:2

Abnormal structure and dysfunction of platelets in CD226 knockout mice
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摘要 目的研究共刺激分子CD226对小鼠血小板功能的调控作用。方法取40周龄老年CD226基因敲除(CD226KO)小鼠为实验组,同龄野生型(WT)C57BL/6小鼠为对照组。取尾静脉血进行血小板计数,剪取小鼠尾尖,检测出血时间;分离小鼠血小板,透射电镜观察血小板超微结构;建立三氯化铁(FeCl_3)诱导的小鼠颈动脉血栓模型,观察CD226KO小鼠与WT小鼠血栓形成的差异。获取人血小板蛋白,通过免疫沉淀和质谱分析,获得与血小板CD226相互作用蛋白的信息。结果与同龄WT小鼠相比,老年CD226KO小鼠血小板数量明显减少,出血时间明显延长。CD226KO小鼠血小板内质网形态发生明显弯曲皱缩变形。在Fe Cl3诱导的小鼠血栓模型中,CD226KO小鼠血栓形成时间明显延长,血栓稳定性显著下降。质谱检测结果提示,人血小板中CD226与脑源性神经生长因子(BDNF)、脂肪酸结合蛋白5(FABP5)、载脂蛋白1(Apo A1)等蛋白具有相互作用。结论 CD226KO小鼠血小板功能障碍,CD226参与血小板生物学活性的发挥。 Objective To study the regulatory effect of co-stimulatory molecule CD226 on platelet function in mice.Methods The 40-week-old CD226 knockout( CD226KO) mice were used as an experimental group and the wild wild-type( WT) C57 BL/6 mice at the same age were designated as a control group. Caudal venous blood was taken for platelet counting. Tail tips of the mice were snipped for the bleeding time measurement. Ultrastructure of platelets was examined by transmission electron microscope. Carotid artery thrombosis model was established by the induction of ferric chloride in mice,to test the difference of platelet function in CD226KO and WT mice. Human platelet protein was harvested for immunoprecipitation( IP) and mass spectrometry analyses to screen the CD226-interactive proteins. Results Aged mice in CD226KO group had significantly lower platelet counts and longer bleeding time compared with the mice in WT group at the same age. Moreover,the scanning electron microscopic image of platelet also indicated that CD226 knockout induced the shrinkage and distortion of platelet endoplasmic reticulum. The Fe Cl3-induced thrombosis model showed that the thrombosis time was significantly longer in CD226KO mice, and thrombus stability was significantly reduced. Mass spectrometry indicated that platelet CD226 interacted with BDNF,FABP5,Apo A1 and other proteins. Conclusion Knockout of CD226 gene significantly affects platelet function in mice,and CD226 molecules are involved in the exertion of biological activity of platelets.
作者 周博 张栋梁 刘雪芹 庄然 张圆 郭树忠 ZHOU Bo;ZHANG Dongliang;LIU Xueqin;ZHUANG Ran;ZHANG Yuan;GUO Shuzhong(Department of Plastic and Reconstructive Surgery,Xijing Hospital;School of Basic Medical Science,Air Force Military Medical University,Xi'an 710032,China)
出处 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2018年第4期309-314,共6页 Chinese Journal of Cellular and Molecular Immunology
基金 国家自然科学基金(81370388) 陕西省重点研发计划(2017SF-214)
关键词 CD226 血小板 基因敲除 CD226 platelet gene knockout
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