miRNA作为血小板储存损伤标志物研究初探*

doi:10.3969/j.issn.1671-2587.2022.02.008

临床输血与检验 ›› 2022, Vol. 24 ›› Issue (2): 176-181.DOI: 10.3969/j.issn.1671-2587.2022.02.008

miRNA作为血小板储存损伤标志物研究初探^*

杨惠宽, 吴千羽, 张嘉仪, 陈小洁, 苑召虎, 魏亚明

510180 广州市第一人民医院输血科,广东省精准输血技术工程研究中心

收稿日期:2021-11-18 发布日期:2022-04-12
通讯作者: 魏亚明,男,主任技师,博士,主要从事干细胞治疗与输血免疫学方面研究,（E-mail）eywym@scut.edu.cn。
作者简介:杨惠宽（1973-）,女,广东广州人,主管技师,本科,主要从事输血免疫方面研究,（E-mail）yhk0209@163.com。并列第一作者：吴千羽（1991-）,女,广东广州人,技师,硕士,主要从事输血免疫学方面研究,（E-mail）584684379@qq.com 。
基金资助:
*本课题受广州市科技计划项目（No.201904010027、202002030427）和广州市临床重大项目（No.2019ZD18）资助

miRNA May Serve as a Promising Marker for Platelet Lesion during Storage in Blood Bank Condition

YANG Hui-kuan, WU Qian-yu, ZHANG Jia-yi, et al

Department of Blood Transfusion,Guangdong Precision Transfusion Technology Engineering Research Center,Guangzhou First People's Hospital,School of Medicine Guangzhou,Guangdong 510180

Received:2021-11-18 Published:2022-04-12

摘要/Abstract

摘要： 目的探讨血小板（PLT）储存过程中microRNA（miRNA）的表达变化,PLT功能相关蛋白表达情况,以及与PLT聚集功能的关系。方法采集机采PLT于（22±2）℃下震荡保存,使用miRNA表达谱芯片检测PLT内miRNA表达及变化。实时荧光定量PCR（qPCR）检测机采PLT储存过程中与聚集功能相关的miRNA表达变化。流式细胞术（FCM）检测PLT凋亡率、PLT内VASP、P2Y12和GPⅡb/Ⅲa。结果对比储存第2天与第5天PLT miRNA表达结果,47种miRNA表达上调,120种下调。储存第8天与第2天相比,28种表达上调,202种下调。变化的miRNA的靶基因主要与PLT脱粒、激活及血小板生长因子（PDGF）受体信号通路有关。其中储存第5天时PLT聚集相关的miR-155、miR-21-5p和miR-21-3p表达下调（P<0.05）,而let-7b、miR-223和miR-3162表达上调（P<0.05）。在（22±2）℃条件下保存的PLT凋亡率随储存天数增加逐步升高。在储存过程中,P2Y12活化程度增高,导致VASP去磷酸化增加,而GPⅡb/Ⅲa含量则在储存的过程中相对稳定。结论大量的miRNA存在于PLT中,且表达量随着PLT保存时间增加而发生变化。这些miRNA的表达变化能影响相应的蛋白水平并最终导致PLT的聚集功能和凋亡发生变化,特定miRNA表达变化具有作为PLT储存损伤的生物学标记可能。

关键词: 血小板, 微小RNA, 储存损伤, 凋亡, GPⅡb/Ⅲa, 血管舒张刺激磷酸蛋白, P2Y12

Abstract: Objective To analyze the expression of miRNAs in stored apheresis platelets and their effects on protein levels associated with platelet aggregation. Methods Healthy male donor platelets were collected and stored at（22±2）℃. The expression profile and changes of miRNA in these platelets were detected by human miRNA array and qRT-PCR. Flow cytometry（FCM）was used to test P2Y12,VASP,GPⅡb/Ⅲa and apoptosis rate in stored platelets. Results Compared the microarray of microRNA expression in platelets stored on the 2nd with 5th days,47 miRNAs expression increased and 120 decreased. Compared the 8th storage day with 2nd day,28 miRNAs increased and 202 miRNAs decreased. Differentially expressed miRNAs were found to be associated with platelet activation,degranulation,PDGF receptor signaling pathway differentiation. As for miRNAs related to platelet aggregation,miR-155,miR-21-5p and miR-21-3p decreased on the 5th day of storage（P<0.05）,while let-7b,miR-223 and miR-3162 increased（P<0.05）. The apoptotic rate of apheresis platelets increased continuously with the increase of storage days. Proteins related to aggregation were also assayed, P2Y12 was activated and then induced the dephosphorylation of VASP. PLT GPⅡb/Ⅲa stayed relatively stable during storage. Conclusion miRNA expressions were found in banked platelets and the expressions changed over time. The changes of miRNA are associated with the corresponding protein level,and eventually lead to the change of platelet aggregation function. Some miRNA expression shows the potential role as biomarkers for platelet lesion during storage in blood bank condition.

Key words: Platelet, MicroRNA, Storage lesion, Apoptosis, GPⅡb/Ⅲa, VASP, P2Y12

中图分类号:

R558

杨惠宽, 吴千羽, 张嘉仪, 陈小洁, 苑召虎, 魏亚明. miRNA作为血小板储存损伤标志物研究初探^*[J]. 临床输血与检验, 2022, 24(2): 176-181.

YANG Hui-kuan, WU Qian-yu, ZHANG Jia-yi, et al. miRNA May Serve as a Promising Marker for Platelet Lesion during Storage in Blood Bank Condition[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2022, 24(2): 176-181.

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miRNA作为血小板储存损伤标志物研究初探^*

miRNA May Serve as a Promising Marker for Platelet Lesion during Storage in Blood Bank Condition

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