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流动条件下新生儿血小板在内皮下的沉积增加:血浆血管性血友病因子的作用

Increased neonatal platelet deposition on subendothelium under flow conditions: the role of plasma von Willebrand factor.

作者信息

Shenkman B, Linder N, Savion N, Tamarin I, Dardik R, Kennet G, German B, Varon D

机构信息

Institute of Thrombosis and Hemostasis, Sheba Medical Center, Tel Hashomer, Israel.

出版信息

Pediatr Res. 1999 Feb;45(2):270-5. doi: 10.1203/00006450-199902000-00019.

Abstract

In vitro platelet function of umbilical cord blood and neonatal peripheral vein blood from full-term newborns was compared with that of adults. Citrated whole blood was subjected to shear stress (1300 s(-1)) on subendothelial extracellular matrix (ECM)-coated wells in a cone and plate(let) analyzer. Adhered platelets on the ECM were quantitated by image analyzer. Both umbilical cord and neonatal peripheral blood platelets demonstrated more extensive adhesion than adult platelets, and similar aggregate formation on ECM. The ability of neonatal platelets to form aggregates on ECM was confirmed by scanning electron microscopy. Similar activation of neonatal and adult platelets after subjection to shear stress, in the suspension phase, was established by flow cytometry, which showed an increase in fibrinogen binding and a decrease in glycoprotein Ib expression on platelet membrane. The difference in adhesion rates between neonatal and adult platelets was preserved even when the hematocrit level of the neonatal blood was adjusted to that of adults. Reconstitution of neonatal or adult platelet-rich plasma with autologous or heterologous red packed cells yielded no change in adhesion and aggregation. When von Willebrand factor-covered plates were used to prevent deposition of plasma von Willebrand factor on the surface, no difference in platelet adhesion was seen between neonatal and adult blood. In conventional aggregometry assay, the response to ristocetin of washed platelets of either neonatal or adult source was higher on addition of plasma from neonates than from adults. Our data suggest that the extensive neonatal platelet deposition on ECM is mediated by plasma von Willebrand factor, which is known to be more multimerized and, therefore, more active in neonates than in adults. This mechanism may provide balanced primary hemostasis in neonates despite the platelet hyporeactivity to agonists without application of shear stress.

摘要

将足月新生儿脐带血和新生儿外周静脉血的体外血小板功能与成年人的进行了比较。在锥板(小)分析仪中,将枸橼酸化全血置于包被有内皮下细胞外基质(ECM)的孔板上,施加剪切应力(1300 s⁻¹)。通过图像分析仪对ECM上黏附的血小板进行定量。脐带血和新生儿外周血的血小板均表现出比成人血小板更广泛的黏附,以及在ECM上形成类似的聚集体。通过扫描电子显微镜证实了新生儿血小板在ECM上形成聚集体的能力。通过流式细胞术确定,在悬浮相中,新生儿和成人血小板在受到剪切应力后有类似的激活,表现为血小板膜上纤维蛋白原结合增加,糖蛋白Ib表达减少。即使将新生儿血液的血细胞比容水平调整至与成人相同,新生儿和成人血小板的黏附率差异仍然存在。用自体或异体红细胞重悬新生儿或成人富含血小板血浆,黏附与聚集无变化。当使用覆盖有血管性血友病因子的板来防止血浆血管性血友病因子沉积在表面时,新生儿和成人血液的血小板黏附无差异。在传统的凝集测定法中,添加新生儿血浆时,新生儿或成人来源的洗涤血小板对瑞斯托霉素的反应比添加成人血浆时更高。我们的数据表明,新生儿血小板在ECM上的广泛沉积是由血浆血管性血友病因子介导的,已知该因子在新生儿中比在成人中多聚化程度更高,因此活性更强。尽管在未施加剪切应力时血小板对激动剂反应低下,但这种机制可能为新生儿提供平衡的初级止血功能。

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