Effect of pathological high shear exposure time on platelet activation and aggregation.

IF 2.1 4区医学 Q3 HEMATOLOGY

Clinical hemorheology and microcirculation Pub Date : 2023-01-01 DOI:10.3233/CH-221567

Tiancong Zhang, Xiaojing Huang, Xuemei Gao, Ling Liu, Dan Chen, Xuanrong Huan, Cui He, Yuan Li

{"title":"Effect of pathological high shear exposure time on platelet activation and aggregation.","authors":"Tiancong Zhang, Xiaojing Huang, Xuemei Gao, Ling Liu, Dan Chen, Xuanrong Huan, Cui He, Yuan Li","doi":"10.3233/CH-221567","DOIUrl":null,"url":null,"abstract":"<p><p>Circulating platelets are sometimes exposed to high shear rate environments due to vascular stenosis, and the effect of transiently elevated pathological high shear rates on platelet activation and aggregation function has not been clarified. The aim of this study was to investigate the effect of pathological high shear rate (8302s-1) exposure time (3.16-25.3 ms) on platelet activation and aggregation function. In addition, by adding active ingredients of antiplatelet drugs such as ASA (an active ingredient of aspirin), Ticagrelor, Tirofiban and GP1BA (platelet membrane protein GPIb inhibitor) in vitro, we studied TXA2, P2Y12-ADP, GPIIb/IIIa-fibrinogen and GPIb /IX/V-vWF receptor pathways to determine platelet activation function mediated by pathological high shear rate. In this study, we designed a set of microfluidic chips with stenosis lengths of 0.5 mm, 1 mm, 2 mm, 3 mm, and 4 mm, all with 80% stenosis, to generate pathological high shear forces that can act at different times. The whole blood flowing through the microchannels was collected by perfusion of sodium citrate anticoagulated whole blood at a physiological arterial shear rate (1500 s-1), and the expression levels of platelet surface activation markers (P-selectin and GP IIb/IIIa) and the degree of platelet aggregation were analyzed by flow cytometry; platelet aggregation patterns were observed by microscopic examination of blood smears. The results showed that shearing significantly increased platelet activation and aggregation levels compared to un-sheared whole blood, and the activation and aggregation levels increased with increasing duration of pathological high shear rate. In vitro inhibition studies showed that ASA barely inhibited the expression of P-selectin and PAC-1 on the platelet surface; Ticagrelor effectively inhibited the expression of both P-selectin and PAC-1; Tirofiban significantly inhibited the expression of PAC-1 on the platelet surface and slightly inhibited the expression of P-selectin; GP1BA significantly inhibited the expression of both. Our results suggest that transient pathological high shear rate (8302s-1) exposure can induce platelet activation in a time-dependent manner; however, the mechanism is more complex and may be due to the following reasons: transient elevated pathological high shear rate activates platelets through the GPIb/IX/V-vWF receptor pathway, and after platelet activation, its surface membrane protein GPIIb/IIIa receptors activate platelets through fibrinogen to form platelet-platelet aggregates, and further activation of active substances such as ADP and TXA2 released by platelet alpha particles, which contribute to the formation of irreversible platelet aggregation.</p>","PeriodicalId":10425,"journal":{"name":"Clinical hemorheology and microcirculation","volume":null,"pages":null},"PeriodicalIF":2.1000,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Clinical hemorheology and microcirculation","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3233/CH-221567","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"HEMATOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Circulating platelets are sometimes exposed to high shear rate environments due to vascular stenosis, and the effect of transiently elevated pathological high shear rates on platelet activation and aggregation function has not been clarified. The aim of this study was to investigate the effect of pathological high shear rate (8302s-1) exposure time (3.16-25.3 ms) on platelet activation and aggregation function. In addition, by adding active ingredients of antiplatelet drugs such as ASA (an active ingredient of aspirin), Ticagrelor, Tirofiban and GP1BA (platelet membrane protein GPIb inhibitor) in vitro, we studied TXA2, P2Y12-ADP, GPIIb/IIIa-fibrinogen and GPIb /IX/V-vWF receptor pathways to determine platelet activation function mediated by pathological high shear rate. In this study, we designed a set of microfluidic chips with stenosis lengths of 0.5 mm, 1 mm, 2 mm, 3 mm, and 4 mm, all with 80% stenosis, to generate pathological high shear forces that can act at different times. The whole blood flowing through the microchannels was collected by perfusion of sodium citrate anticoagulated whole blood at a physiological arterial shear rate (1500 s-1), and the expression levels of platelet surface activation markers (P-selectin and GP IIb/IIIa) and the degree of platelet aggregation were analyzed by flow cytometry; platelet aggregation patterns were observed by microscopic examination of blood smears. The results showed that shearing significantly increased platelet activation and aggregation levels compared to un-sheared whole blood, and the activation and aggregation levels increased with increasing duration of pathological high shear rate. In vitro inhibition studies showed that ASA barely inhibited the expression of P-selectin and PAC-1 on the platelet surface; Ticagrelor effectively inhibited the expression of both P-selectin and PAC-1; Tirofiban significantly inhibited the expression of PAC-1 on the platelet surface and slightly inhibited the expression of P-selectin; GP1BA significantly inhibited the expression of both. Our results suggest that transient pathological high shear rate (8302s-1) exposure can induce platelet activation in a time-dependent manner; however, the mechanism is more complex and may be due to the following reasons: transient elevated pathological high shear rate activates platelets through the GPIb/IX/V-vWF receptor pathway, and after platelet activation, its surface membrane protein GPIIb/IIIa receptors activate platelets through fibrinogen to form platelet-platelet aggregates, and further activation of active substances such as ADP and TXA2 released by platelet alpha particles, which contribute to the formation of irreversible platelet aggregation.

查看原文本刊更多论文

病理性高剪切暴露时间对血小板活化和聚集的影响。

由于血管狭窄，循环血小板有时暴露于高剪切率环境中，而病理性高剪切率的短暂升高对血小板活化和聚集功能的影响尚不清楚。本研究旨在探讨病理性高剪切速率(8302s-1)暴露时间(3.16 ~ 25.3 ms)对血小板活化和聚集功能的影响。此外，我们通过体外添加ASA(阿司匹林的有效成分)、替格瑞洛、替罗非班、GP1BA(血小板膜蛋白GPIb抑制剂)等抗血小板药物的有效成分，研究TXA2、P2Y12-ADP、GPIIb/ iiia -纤维蛋白原、GPIb /IX/V-vWF受体途径，测定病理性高剪切率介导的血小板活化功能。在本研究中，我们设计了一组狭窄长度分别为0.5 mm、1mm、2mm、3mm和4mm的微流控芯片，狭窄程度均为80%，以产生可在不同时间起作用的病状高剪切力。以生理动脉剪切速率(1500 s-1)灌注柠檬酸钠抗凝全血采集微通道内的全血，流式细胞术分析血小板表面活化标志物(p -选择素、GP IIb/IIIa)表达水平及血小板聚集程度;血液涂片镜检观察血小板聚集模式。结果表明，与未剪切全血相比，剪切后血小板活化和聚集水平显著升高，且活化和聚集水平随病理性高剪切率持续时间的延长而升高。体外抑制实验表明，ASA对血小板表面p -选择素和PAC-1的表达几乎没有抑制作用;替格瑞洛有效抑制p -选择素和PAC-1的表达;替罗非班显著抑制血小板表面PAC-1的表达，轻微抑制p -选择素的表达;GP1BA显著抑制两者的表达。我们的研究结果表明，短暂的病理性高剪切速率(8302s-1)暴露可以以时间依赖性的方式诱导血小板活化;然而，其机制更为复杂，可能是由于以下原因:一过性升高的病理性高剪切速率通过GPIb/IX/V-vWF受体途径激活血小板，血小板活化后，其表面膜蛋白GPIIb/IIIa受体通过纤维蛋白原激活血小板形成血小板-血小板聚集，并进一步激活血小板α粒子释放的ADP、TXA2等活性物质，形成不可逆的血小板聚集。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Clinical hemorheology and microcirculation 医学-外周血管病

CiteScore

4.30

自引率

33.30%

发文量

170

期刊介绍： Clinical Hemorheology and Microcirculation, a peer-reviewed international scientific journal, serves as an aid to understanding the flow properties of blood and the relationship to normal and abnormal physiology. The rapidly expanding science of hemorheology concerns blood, its components and the blood vessels with which blood interacts. It includes perihemorheology, i.e., the rheology of fluid and structures in the perivascular and interstitial spaces as well as the lymphatic system. The clinical aspects include pathogenesis, symptomatology and diagnostic methods, and the fields of prophylaxis and therapy in all branches of medicine and surgery, pharmacology and drug research. The endeavour of the Editors-in-Chief and publishers of Clinical Hemorheology and Microcirculation is to bring together contributions from those working in various fields related to blood flow all over the world. The editors of Clinical Hemorheology and Microcirculation are from those countries in Europe, Asia, Australia and America where appreciable work in clinical hemorheology and microcirculation is being carried out. Each editor takes responsibility to decide on the acceptance of a manuscript. He is required to have the manuscript appraised by two referees and may be one of them himself. The executive editorial office, to which the manuscripts have been submitted, is responsible for rapid handling of the reviewing process. Clinical Hemorheology and Microcirculation accepts original papers, brief communications, mini-reports and letters to the Editors-in-Chief. Review articles, providing general views and new insights into related subjects, are regularly invited by the Editors-in-Chief. Proceedings of international and national conferences on clinical hemorheology (in original form or as abstracts) complete the range of editorial features.