天津医药

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高密度脂蛋白对血小板α颗粒膜糖蛋白及CD63 表达的影响

杨升华1,杜大勇2,柳杨3,王双双4,李运田5   

  1. 1. 广州市， 南方医科大学研究生院
    2. 中国人民解放军第305医院心脏中心二病区
    3. 中国人民解放军第305医院心脏中心暨全军冠心病诊疗中心
    4. 中国人民解放军第305医院中心实验室生化室
    5. 中国人民解放军第305医院心脏中心、总参谋部心血管中心暨全军冠心病诊疗中心心二病区
  • 收稿日期:2012-11-08 修回日期:2013-03-01 出版日期:2013-07-15 发布日期:2013-07-15
  • 通讯作者: 杨升华

Effects of High Density Lipoprotein on Expression of Alpha-Granulemembrane Protein (CD62P) and Lysosome Intact Membrane Protein (CD63) on Platelet

YANG Sheng hua1,DU Dayong 2,LIU Yang 2,WANG Shuangshuang 2,LI Yuntian 2   

  1. 1. Graduate School of Southern Medical University， Guangzhou 510515， China
    2. Heart Center of Chinese No. 305 Hospital of People’ s Liberation Army
  • Received:2012-11-08 Revised:2013-03-01 Published:2013-07-15 Online:2013-07-15
  • Contact: YANG Sheng hua

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摘要： 【摘要】 目的  研究高密度脂蛋白(HDL)对凝血酶刺激的人血小板α颗粒膜糖蛋白(CD62P或P-选择素)、溶酶体膜糖蛋白(CD63)表达的影响。方法  3组等体积手工制备的洗涤血小板分别加入0.5、1和10U/mL的凝血酶(低、中、高凝血酶组),另外3组同样含量的凝血酶加入37℃孵育15min的HDL,分别为低、中、高复合组。同时37℃环境下孵育10min,以流式细胞仪检测各组CD62P、CD63的表达。结果  低、中、高复合组CD62P表达率分别为11.55%±1.34%,17.19%±0.17%,19.79%±0.32%,均低于相应凝血酶组的18.14%±1.50%,26.24%±0.77%,80.38%±5.66%(均P<0.01),低、中、高复合组CD63表达也低于相应的凝血酶组即(2.92%±0.22%、4.20%±0.98%,5.12%±0.09%vs8.09%±0.48%、14.15%±1.39%、24.48%±1.71%,均P<0.01)。低、中复合组血小板CD62P、CD63抑制率均低于高复合组(均P<0.05)。结论  在体外,HDL抑制了凝血酶刺激的血小板α颗粒膜糖蛋白和溶酶体膜糖蛋白的表达。

关键词: 脂蛋白类, HDL, P-选择素, 凝血酶, 流式细胞仪, 溶酶体相关膜蛋白质 1, 抗原, CD36

Abstract: Abstact   Objective   To investigate the effect of High Density Lipoprotein on expression of CD62P and CD63 on thrombin-activated platelet in vitro. Methods   Equivalent volume of washed platelet prepared by hand was preincubated with HDL(1mg/ml) in 37℃ water for 15 minutes ，then stimulated with different concentrations of thrombin(0.5U/ml,1U/ml,10U/ml) for 10 minutes in water of 37℃.Meanwhile another three groups of washed platelet was incubated with thrombin(0.5U/ml,1U/ml,10U/ml) for 10minutes,respectively.The CD62P and CD63 from each sample were analyzed by flow cytometry(FCM). Results   The CD62P positive rate of HDL-preincubated groups were significantly lower than the groups of different concentrations of thrombin(0.5U/ml,1U/ml,10U/ml) in the absence of HDL, 11.55%±1.34% vs 18.14%±1.50%,17.19%±0.17% vs 26.24%±0.77%,19.79%±0.32% vs 80.38%±5.66%;P value <0.001.Meanwhile, The CD63 positive rate of HDL-preincubated groups were also significantly lower the thrombin-treated (0.5U/ml,1U/ml,10U/ml) groups without HDL,namely,8.09%±0.48% vs 2.92%±0.22%(P<0.001),14.15%±1.39% vs 4.20%±0.98%(P<0.001),24.48%±1.71% vs 5.12%±0.09%(P<0.001). Conclusion   HDL inhibits expression of alpha-granulemembrane protein (CD62P) and lysosome intact membrane protein(CD63) on thrombin-stimulated platelet in vitro.

Key words: poproteins, HDL, P-selectin, thrombin, flow cytometry, lysosomal-associated membrane protein 1, antigens, CD36