The shear rate promotes pinocytosis of extracellular dextran in platelets.

Masataka Inoue, Masahiro Ohwada, Nobuo Watanabe
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Abstract

Background: Several conventional studies focused on platelet pinocytosis for possible utilization as drug delivery systems. Although platelet pinocytosis is important in such utilization, the impact of the shear rate on pinocytosis is unclear.

Objective: Our objective was to investigate the relationship between shear rate and platelet pinocytosis in vitro. In addition, this study addressed the change in platelet aggregation reactivity with adenosine diphosphate (ADP) stimulation after pinocytosis.

Method: Porcine platelet-rich plasma was mixed with fluorescein isothiocyanate (FITC)-conjugated dextran and incubated for 15 min under shear conditions of 0, 500, and 1500 s-1. After incubation, confocal microscopic scanning and three-dimensional rendering were performed to confirm the internalization of FITC-dextran into platelets. The amount of FITC-dextran accumulated via platelet pinocytosis was compared using flow cytometry at each shear rate. In addition, light transmission aggregometry by ADP stimulation was applied to platelets after pinocytosis.

Results: The amount of intracellular FITC-dextran increased with higher shear rates. Platelets with increased amounts of intracellular FITC-dextran did not show changes in the aggregation reactivity to ADP.

Conclusions: A higher shear rate promotes platelet pinocytosis, but enhanced pinocytosis does not affect aggregation sensitivity, which is stimulated by ADP.

剪切率可促进血小板中细胞外葡聚糖的针吞作用。
背景:一些传统研究关注血小板的纤溶作用,以便将其用作药物输送系统。虽然血小板的蛲吞作用在这种利用中很重要,但剪切率对蛲吞作用的影响尚不清楚:我们的目的是研究剪切率与体外血小板蛲虫吞噬作用之间的关系。此外,本研究还探讨了血小板在二磷酸腺苷(ADP)刺激下的聚集反应性在针吞后的变化:方法:将猪血小板丰富血浆与异硫氰酸荧光素(FITC)共轭葡聚糖混合,在 0、500 和 1500 s-1 的剪切条件下孵育 15 分钟。孵育后,进行共聚焦显微镜扫描和三维渲染,以确认 FITC 葡聚糖是否内化到血小板中。在每种剪切速率下,使用流式细胞术比较了通过血小板针吞积累的 FITC-葡聚糖的数量。此外,通过 ADP 刺激对血小板针吞后进行透光聚集测定:结果:细胞内 FITC-葡聚糖的含量随着剪切率的升高而增加。细胞内 FITC-葡聚糖含量增加的血小板对 ADP 的聚集反应性没有变化:结论:较高的剪切率可促进血小板的针形细胞增多,但针形细胞增多并不影响聚集敏感性,聚集敏感性受到 ADP 的刺激。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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