显微三维成像技术分析替罗非班抑制血小板黏附聚集的量效关系

引用本文: 何菲, 张天聪, 陈丹, 丁玲, 邓素容, 米永华, 刘占术, 李远. 显微三维成像技术分析替罗非班抑制血小板黏附聚集的量效关系[J]. 分析化学, 2022, 50(2): 244-252. doi: 10.19756/j.issn.0253-3820.210671 shu

Citation: HE Fei, ZHANG Tian-Cong, CHEN Dan, DING Ling, Deng Su-Rong, MI Yong-Hua, LIU Zhan-Shu, LI Yuan. Analysis of Dose-effect Relationship of Tirofiban Inhibiting Platelet Adhesion Aggregation Using Microscopic Three-dimensional Imaging Technology[J]. Chinese Journal of Analytical Chemistry, 2022, 50(2): 244-252. doi: 10.19756/j.issn.0253-3820.210671 shu

显微三维成像技术分析替罗非班抑制血小板黏附聚集的量效关系

何菲 ^1, ,
张天聪 ^1, ,
陈丹 ^1, ,
丁玲 ^2, ,
邓素容 ^1, ,
米永华 ^1, ,
刘占术 ^1, ,
李远 ^{1,
,}

1.
重庆医科大学附属永川医院中心实验室,永川 402160;
2.
重庆市血液中心永川分中心,永川 402160

通讯作者: 李远,E-mail:liyuan_1985999@163.com

基金项目:
国家自然科学基金项目（No.11702047）、重庆市社会事业与民生保障科技创新专项项目（No.cstc2017shmsA130009）、重庆市医学科研计划项目（No.2017MSXM079）、永川区社会民生科技创新专项（No.Ycstc2018cc0206）和重庆医科大学附属永川医院院级课题（No.YJLC202033）资助

摘要: 建立了一种显微三维成像技术用于定性和定量评价替罗非班(血小板膜糖蛋白(GP) Ⅱb/Ⅲa受体拮抗剂)抑制血小板黏附聚集的量效关系。在健康志愿者富血小板血浆(PRP)中加入0~50 μg/mL替罗非班进行处理，将处理后的PRP分别滴加在胶原纤维蛋白和玻璃表面上，诱导血小板活化聚集，通过激光显微三维成像技术观察不同条件下血小板聚集体的显微三维形态学图像，并量化血小板聚集体的体积、面积覆盖率、血小板黏附数量和活化血小板数量比例。结果表明，利用激光显微三维成像技术可直观地观察到胶原纤维蛋白和玻璃表面形成的血小板聚集体的三维显微形态。胶原纤维蛋白表面形成的血小板聚集体呈薄片状，玻璃表面形成的血小板聚集体呈山丘状。对于胶原纤维蛋白表面，替罗非班终浓度为1.56~12.5 μg/mL时，随其浓度增加，血小板聚集体体积和面积覆盖率显著降低；当替罗非班浓度增加至25和50 μg/mL，仅见血小板在胶原纤维蛋白上发生黏附，血小板黏附数量和活化血小板比例随浓度增加而显著降低。在玻璃表面，经1.56 μg/mL替罗非班处理，血小板聚集体体积和面积覆盖率降低；3.15~50 μg/mL替罗非班可完全抑制血小板聚集，并且血小板黏附数量和活化血小板比例随替罗非班浓度增加而显著降低。综上，3.15~12.5 μg/mL替罗非班既可完全抑制血小板在玻璃表面的聚集，又保留了部分血小板在胶原纤维蛋白表面上的聚集能力。采用显微三维成像技术定性和定量分析替罗非班抑制血小板在胶原纤维蛋白和玻璃表面的黏附聚集的量效关系，可为临床合理选择替罗非班的剂量提供新的分析方法。

关键词:

English

Analysis of Dose-effect Relationship of Tirofiban Inhibiting Platelet Adhesion Aggregation Using Microscopic Three-dimensional Imaging Technology

1.
Central Laboratory of Yongchuan Hospital, Chongqing Medical University, Yongchuan 402160, China;
2.
Yongchuan Sub-center of Chongqing Blood Center, Yongchuan 402160, China

Corresponding author: LI Yuan, liyuan_1985999@163.com

Received Date: 11 August 2021
Revised Date: 08 December 2021
Fund Project:
Supported by the National Natural Science Foundation of China (No.11702047), the Special Project of Science and Technology Innovation for People′s Livelihood Security of Chongqing (No.cstc2017shmsA130009), the Chongqing Medical Research Program (No.2017MSXM079), the Special Project of Science and Technology Innovation for Livelihood Security of Yongchuan (No.Ycstc2018cc0206) and the Research Project of Yongchuan Hospital, Chongqing Medical University(No.YJLC202033)

Abstract: The dose-effect relationship of tirofiban (a platelet membrane glycoprotein (GP) Ⅱb/Ⅲa receptor antagonist) in inhibiting platelet adhesion and aggregation was qualitatively and quantitatively evaluated using a novel microscopic three-dimensional (3D) imaging technology. The platelet-rich plasma (PRP) of healthy volunteers was firstly treated by 0-50 μg/mL tirofiban, and then added to collagen fibrin and glass surfaces respectively to induce platelet activation and aggregation. The 3D morphological images of platelet adhesion and aggregation under different experimental conditions were observed by laser microscopic 3D imaging technology. The volume and area coverage rate of platelet aggregates, and the number and activated count of adhesion platelet were quantified. The results showed that the 3D micro-morphology of platelet aggregates formed on the surfaces of collagen fibrin and glass could be visually observed by the laser microscopic 3D imaging technology. The platelet aggregates formed on the surface of collagen fibrin were thin, and those on the surface of glass were hill-like. When the final concentration of tirofiban was 1.56-12.5 μg/mL, the volume and area coverage rate of platelet aggregate decreased significantly with the increase of tirofiban final concentration. When the concentration increased to 25 and 50 μg/mL, platelets adhered to collagen fibrin only, and the number of platelet adhesion and the proportion of activated platelets decreased significantly with the increase of concentration. On the glass surface, the tirofiban with concentration of 1.56 μg/mL could reduce the volume and area coverage rate of platelet aggregates; and when the concentration was 3.15-50 μg/mL, it could completely inhibit platelet aggregation, and the number of platelet adhesion and the proportion of activated platelets decreased with the increase of tirofiban concentration. As a result, when the concentration was 3.15-12.5 μg/mL, tirofiban could not only completely inhibit platelet aggregation on the glass surface but retain partial aggregation ability on collagen fibrin surface. The microscopic 3D imaging technique could quantitatively and qualitatively analyze the dose-effect relationship of tirofiban inhibiting platelet adhesion aggregation on collagen fibrin surface and glass surface, which provided a new analytical method for rational clinical selection of tirofiban dose.

Key words:

Platelet aggregation
/ Microscopic three-dimensional morphology
/ Collagen fiber
/ Glass
/ Tirofiban
/ Does-effective relationship

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

显微三维成像技术分析替罗非班抑制血小板黏附聚集的量效关系

通讯作者: 李远,E-mail:liyuan_1985999@163.com

English

Analysis of Dose-effect Relationship of Tirofiban Inhibiting Platelet Adhesion Aggregation Using Microscopic Three-dimensional Imaging Technology

Corresponding author: LI Yuan, liyuan_1985999@163.com

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

显微三维成像技术分析替罗非班抑制血小板黏附聚集的量效关系

通讯作者: 李远,E-mail:liyuan_1985999@163.com

English

Analysis of Dose-effect Relationship of Tirofiban Inhibiting Platelet Adhesion Aggregation Using Microscopic Three-dimensional Imaging Technology

Corresponding author: LI Yuan, liyuan_1985999@163.com

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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