应用激光扫描共聚焦显微术观察血小板活化及其形态改变  

The Detection of Platelet Activation and its Morphological Change by Laser Scanning Confocal Microscopy

作　　者：刘嵘[1] 魏文宁[1] 

机构地区：[1]华中科技大学同济医学院协和医院血液病研究所,武汉430022

出　　处：《血栓与止血学》2006年第5期197-200,共4页Chinese Journal of Thrombosis and Hemostasis

基　　金：湖北省科技攻关计划项目(No.2004AA301C32)卫生部临床学科重点资助项目(No.2004-468)

摘　　要：目的建立应用激光扫描共聚焦显微术(laser scanning confocal microscopy,LSCM)观察血小板活化及其形态改变的方法。方法用不同浓度的胶原诱导血小板活化,通过LSCM观察与特异性荧光(FITC/PE)抗体结合的血小板膜糖蛋白Ⅱb/Ⅲa(GPⅡb/Ⅲa)荧光强度的变化,判断血小板微颗粒(platelet microparticle,PMP)的产生,同时观察正常人和糖尿病视网膜病变患者血小板形态的差异。结果加入诱导剂后血小板活化,镜下观察特异性抗体标记的血小板荧光强度呈规律性变化。在一定浓度时诱导剂胶原的加入可引起血小板GPⅡb/Ⅲa表达量的变化,且随着胶原浓度的增加,GPⅡb/Ⅲa表达量也有增加的趋势。并观察到糖尿病视网膜病变患者血小板有伪足伸出,末端附近有释放的微颗粒环绕,与正常人血小板形态有明显差异。结论应用LSCM可观察到诱导剂致血小板活化引起GPⅡb/Ⅲa表达量的变化以及血小板微颗粒的释放,同时可认为血小板形态改变的观察在血栓病血小板活化程度的判断上具有一定的价值。Objective To establish the method of detecting platelet activation and morphological change directly by using the laser scanning confocal microscopy. Methods We induced platelet to activate with different concentration collagen, detected the fluorescence intensity change of platelet membrance glycoprotein （GP） Ⅱb/Ⅲa complex combining specific fluorescence （FITC/PE） antibody and judged the release of platelet microparticles, observed the difference of platelet morphology between the normal people and diabetic retinopathy by laser scanning confocal microscopy Results Platelets were activated by inducer. We found the fluorescence intensity of the platelets labeled specific antibody concentrations, the addition of collagen induced the change of GP Ⅱb/ glycoprotein GP Ⅱb/Ⅲa complex expression levels show increased tende changed in regular way. At certain Ⅲ a complex expression levels. And ncy with the growth of collagen concentration. There were presence of pseudopodia and microparticles surrounding the platelet body or be near the terminal ends in diabetic retinopathy. They were different in platelet morphology as compared with normal people ）. Conclusion We can detect platelet membrance glycoprotein GP Ⅱb/Ⅲa complex expression levels change because of platelets activation induced by collagen and release of platelet microparticles by using the laser scanning confocal microscopy. Meanwhile it is inferred observation of platelets morphology change have some value in diagnosis of platelets activation levels in thrombosis.

关 键 词：激光扫描共聚焦显微术 血小板微颗粒 糖尿病视网膜病变 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]