Liu Shuchuan, Wei Li, Zhang Yan, Xu Mengyuan, Wang Chen, Zhou Jin
Department of Hematology, the First Affiliated Hospital, Harbin Medical University, Harbin 150001, China.
Department of Cardiology, the Fourth Affiliated Hospital, Harbin Medical University, Harbin 150001, China.
Thromb Res. 2014 Apr;133(4):645-51. doi: 10.1016/j.thromres.2013.12.043. Epub 2014 Jan 6.
Amniotic fluid contains various procoagulant factors involved in intro-vascular amniotic fluid-induced coagulopathies. During the progression of normal pregnancy, microparticles would be shed off from cells and accumulate in amniotic fluid over time. In this study, our aims were to investigate the cellular origin and procoagulant entity of these microparticles.
Twenty amniotic fluid samples from healthy parturient women were collected, and the microparticles were isolated and stained with phycoerythrin-labeled antibodies to CD138, CD41a, CD144 and CD11b to identify their cellular origin. Their phosphatidylserine and tissue factor expression levels were quantified with fluorescein isothiocyanate-labeled annexin V and anti-tissue factor antibody staining. Their procoagulant activity was tested with plasma coagulation assay and factor Xase and prothrombinase assays.
Phenotypic analysis showed 36.8% and 33.8% of amniotic fluid microparticles positive for CD138 and CD11b, respectively, indicating their epithelial cell or leukocyte origin. Of these microparticles, 66.3±5.9% expressed phosphatidylserine while 37.4±4.1% expressed tissue factor. In addition, amniotic fluid microparticles could significantly shorten the plasma coagulation time and increase the production of factor Xa and thrombin. Inhibition assays with annexin V and anti-tissue factor antibody confirmed the coagulation effects of amniotic fluid microparticles.
The microparticles derived from epithelial and leukocytes may be a mechanism of amniotic fluid-induced coagulopathies.
羊水含有多种促凝血因子，参与血管内羊水诱导的凝血病。在正常妊娠过程中，微粒会从细胞上脱落，并随时间在羊水中积累。在本研究中，我们的目的是研究这些微粒的细胞来源和促凝血实体。
收集20例健康产妇的羊水样本，分离微粒，并用藻红蛋白标记的抗CD138、CD41a、CD144和CD11b抗体进行染色，以确定其细胞来源。用异硫氰酸荧光素标记的膜联蛋白V和抗组织因子抗体染色定量其磷脂酰丝氨酸和组织因子表达水平。用血浆凝固试验、因子X酶和凝血酶原酶试验检测其促凝血活性。
表型分析显示，分别有36.8%和33.8%的羊水微粒CD138和CD11b呈阳性，表明其来源于上皮细胞或白细胞。在这些微粒中，66.3±5.9%表达磷脂酰丝氨酸，而37.4±4.1%表达组织因子。此外，羊水微粒可显著缩短血浆凝固时间，并增加因子Xa和凝血酶的产生。用膜联蛋白V和抗组织因子抗体进行的抑制试验证实了羊水微粒的凝血作用。
来源于上皮细胞和白细胞的微粒可能是羊水诱导凝血病的一种机制。
