体外组织因子微颗粒的表达及其促凝活性的分析

摘要

目的：体外细胞系培养刺激获得组织因子微颗粒（TF+MP）并对其促凝活性进行研究。方法用不同浓度脂多糖（LPS）刺激人急性单核细胞白血病细胞系（THP-1），梯度离心后重悬分离获得微颗粒（MP）；在正常人去 MP 血浆中加入 MP，采用凝血酶生成试验分别检测组织因子（TF）和磷酯酰丝氨酸（PS）的凝血酶生成能力；采用 ZYMUPHEN MP-TF 试剂盒检测 TF 阳性的 MP 中 TF 的促凝活性，选用0.45和0.65μm 的滤膜滤过 MP，分别检测直径<0.45和<0.65μm 的 MP 颗粒的促凝活性。结果凝血酶生成试验和发色底物功能检测结果显示体外表达的 MP 具有促凝活性。凝血酶生成试验结果表明 MP 的凝血活性与 LPS 的刺激呈浓度依赖性增强。直径>0.65μm 的 MP 颗粒具有较强的促凝活性。结论体外经 THP-1单核细胞系培养表达 MP具有一定的促凝活性。%Objective To study the procoagulant property of tissue factor-positive microparticles(TF+MP) generated in vitro through the culture and stimulation of cell line. Methods Human acute monocytic leukemia cell line(THP-1)was stimulated by different concentrations of lipopolysaccharide(LPS). Microparticle(MP)was separated by gradient centrifugation. Thrombin generation abilities of tissue factor(TF)and phosphatidylserine (PS)were determined by thrombin generation test by adding MP into MP-depleted plasma. ZYMUPHEN MP-TF kit was performed to determine TF procoagulant property. The 0.45 and 0.65 μm filters were selected to determine MP procoagulant properties(< 0.45 and < 0.65 μm). Results Both thrombin generation test and chromogenic substrate test demonstrated a procoagulant property of MP generated in vitro. Thrombin generation test further showed that the procoagulant property of MP increased by the addition of LPS in a concentration-dependent manner. MP > 0.65 μm had a strong procoagulant property. Conclusions MP generated in vitro through the culture and stimulation of THP-1 demonstrates certain procoagulant property.