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使用流式细胞术对猫血小板中血管舒张刺激磷蛋白磷酸化进行定量分析来评估氯吡格雷对P2Y的抑制作用。

Assessment of P2Y Inhibition by Clopidogrel in Feline Platelets Using Flow Cytometry Quantification of Vasodilator-Stimulated Phosphoprotein Phosphorylation.

作者信息

Li Ronald H L, Nguyen Nghi, Rosati Tommaso, Jandrey Karl

机构信息

Department of Veterinary Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, Davis, CA, United States.

William R. Pritchard Veterinary Medical Teaching Hospital, University of California, Davis, Davis, CA, United States.

出版信息

Front Vet Sci. 2020 May 27;7:267. doi: 10.3389/fvets.2020.00267. eCollection 2020.

Abstract

The primary objective of this study was to evaluate a novel flow cytometry-based assay of quantifying platelet phosphorylation of vasodilator-stimulated phosphoprotein (P-VASP) in cats that received clopidogrel treatment. Eight healthy cats received 18.75 mg PO q24h of clopidogrel for 7 days. Prior to and after clopidogrel treatment, blood was collected for ADP-induced light transmission aggregometry (LTA) and P-VASP measurement by flow cytometry. Flow cytometry measurement of P-VASP levels was used to derive platelet reactivity index (PRI) before and after clopidogrel treatment. Based on P-VASP and LTA findings, platelet response to ADP was significantly attenuated after 7 days of clopidogrel treatment. By eliciting the competing platelet pathways of P2Y and cAMP using ADP and PGE, respectively, ADP had no effect on P-VASP levels following clopidogrel treatment ( = 0.94). Clopidogrel also significantly decreased PRI from 28.84 ± 28.52% to 1.69 ± 12.39% ( = 0.0078). PRI on day 8 correlated moderately with the degree of slope inhibition on LTA ( = -0.4, = 0.4). Flow cytometry analysis of P-VASP is effective at monitoring the inhibitory effects of clopidogrel on feline platelets.

摘要

本研究的主要目的是评估一种基于流式细胞术的新方法,用于量化接受氯吡格雷治疗的猫体内血管舒张刺激磷蛋白(P-VASP)的血小板磷酸化水平。八只健康猫每天口服18.75 mg氯吡格雷,每24小时一次,共7天。在氯吡格雷治疗前后,采集血液用于ADP诱导的光透射聚集试验(LTA)以及通过流式细胞术测量P-VASP。通过流式细胞术测量P-VASP水平来得出氯吡格雷治疗前后的血小板反应性指数(PRI)。基于P-VASP和LTA的结果,氯吡格雷治疗7天后,血小板对ADP的反应明显减弱。分别使用ADP和PGE激发P2Y和cAMP的竞争性血小板途径后,氯吡格雷治疗后ADP对P-VASP水平无影响(P = 0.94)。氯吡格雷还使PRI从28.84±28.52%显著降低至1.69±12.39%(P = 0.0078)。第8天的PRI与LTA上的斜率抑制程度呈中度相关(r = -0.4,P = 0.4)。P-VASP的流式细胞术分析可有效监测氯吡格雷对猫血小板的抑制作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/003c/7267589/139ad2a0a301/fvets-07-00267-g0001.jpg

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