血小板膜受体检测方法的建立及细胞因子的作用研究

Study on Detecting Membrane Receptor of Platelets by Flow Cytometry(FCM)and Effect of Cell Factor

目的建立流式细胞术检测血小板膜蛋白GMP-140的实验方法,分析血小板生成素(TPO)与GM-CSF因子对血小板膜糖蛋白GMP-140表达的影响。方法以生理盐水、TPO溶液及GM-CSF溶液为对照和实验条件与血小板孵育,加入荧光标记的单克隆抗体,应用流式细胞术检测血小板膜蛋白GMP-140的表达率。结果125ng/mlTPO实验组GMP-140表达率与对照组相比有显著性差别(P<0.001);100ng/mlGM-CSF实验组与对照组相比无显著性差别(P>0.5)。结论成功建立了流式细胞术检测血小板膜蛋白GMP-140的实验方法。TPO在一定浓度下对血小板有刺激激活作用,100ng/ml浓度的GM-CSF在体外直接作用于血小板对其无明显刺激作用。

Objective To establish the method of detecting platelets membrane protein GMP-140 by FCM,and to analyse the expressive effect of GMP-140 after by acting cell factor. Methods Salt water,TPO and GM-CSF were incubated wit hplates in room tempreture and divided into control group and experiment group. The expression of plate CMP-140 was determined by FCM through adding antibodies conjugating fluorescence. Results The expression of plate GMP-140 was significantly higher in 125ng/ml TPO group compared with that of salt water control group（P〈0. 001）,and there was no statistical diffenrence between 100 ng/ml GM-CSF group and control group（P〉0.5）. Conclusion The FCM method to detect plate GMP-140 is success. 125 ng/ml TPO could activate plates and 100 ng/ml GM-CSF couldn＇t in vitro.