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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 investigate the effect of TF + MP on the procoagulant activity in vitro. Methods Human acute monocytic leukemia cell line (THP-1) was stimulated with different concentrations of lipopolysaccharide (LPS), and then resuspended to obtain micro-particles (MP) after gradient centrifugation. MP was added to normal MP plasma and thrombin The generation test was used to detect the thrombin generation ability of tissue factor (TF) and phosphatidylserine (PS) respectively. The procoagulant activity of TF in TF-positive MP was detected by ZYMUPHEN MP-TF kit. Filters of 0.45 and 0.65 μm MP was filtered to detect the procoagulant activity of MP particles with diameters <0.45 and <0.65 μm, respectively. Results The results of thrombin generation test and chromogenic substrate function test showed that MP expressed in vitro had pro-coagulant activity. Thrombin generation test results showed that the coagulation activity of MP was stimulated in a concentration-dependent manner. MP particles> 0.65μm in diameter have strong procoagulant activity. Conclusions The expression of MP in THP-1 monocyte cell line in vitro has some procoagulant activity.