Holme P A, Solum N O, Brosstad F, Røger M, Abdelnoor M
Research Institute for Internal Medicine, Rikshospitalet, Norway.
Thromb Haemost. 1994 Nov;72(5):666-71.
Platelet vesiculation in vitro is correlated to platelet activation. It was therefore considered of interest to see if microvesicles (MV) are present in blood in clinical situations associated with platelet activation in vivo. Patients with both activated coagulation and fibrinolysis, implying that thrombin has been generated, suit such a purpose. Thus, the aim of this study was to investigate whether microvesicles could be detected in patients with activated coagulation and fibrinolysis, as diagnosed by the presence of soluble fibrin (positive ethanol gelation tests) and positive tests for fibrin degradation products (FDP). Platelet-rich plasma was prepared from citrated blood from patients (n = 22) and healthy controls (n = 32) matched as to age and sex. The intact platelets were removed from plasma by centrifugation. Any MV present were isolated from the platelet-free plasma by a filtration procedure, washed, solubilized in Triton X-100 and subjected to SDS-PAGE with Western blotting using a MAb against GPIIb alpha as an indicator of the presence of microvesicles. All of the 22 patients showed the presence of microvesicles detectable by the content of GPIIb alpha, whereas this could be observed in only 4 out of the 32 normal controls and then in small or trace amounts only. The presence of microvesicles among cell-derived material in the plasma of two of the patients was also confirmed by electron microscopy. To the best of our knowledge this is the first report on the presence of microvesicles in plasma from patients with both activated coagulation and fibrinolysis.(ABSTRACT TRUNCATED AT 250 WORDS)
血小板体外囊泡化与血小板活化相关。因此，研究在体内与血小板活化相关的临床情况下血液中是否存在微泡（MV）具有重要意义。同时存在凝血和纤溶激活，这意味着已生成凝血酶的患者符合这一研究目的。因此，本研究旨在调查在经可溶性纤维蛋白（乙醇凝胶试验阳性）和纤维蛋白降解产物（FDP）检测呈阳性诊断为凝血和纤溶激活的患者中是否能检测到微泡。从年龄和性别匹配的患者（n = 22）和健康对照者（n = 32）的枸橼酸盐血中制备富血小板血浆。通过离心从血浆中去除完整血小板。通过过滤程序从无血小板血浆中分离出任何存在的微泡，洗涤后，用 Triton X - 100溶解，并使用抗 GPIIbα单克隆抗体进行 SDS - PAGE和蛋白质印迹，以指示微泡的存在。22例患者均显示可通过 GPIIbα含量检测到微泡，而32例正常对照者中只有4例能观察到微泡，且含量很少或仅为微量。两名患者血浆中细胞源性物质中微泡的存在也通过电子显微镜得到证实。据我们所知，这是关于同时存在凝血和纤溶激活的患者血浆中存在微泡的首次报道。（摘要截短于250字）
