基于非标记定量蛋白组学对原发性血小板增多症患者血小板活化相关差异蛋白的分析  

Analysis of Differential Proteins Related to Platelet Activation in Patients with Essential Thrombocythemia Based on Label-Free Quantitative Technology

作　　者：李芋锦 马菊宁 王子卿 杨二鹏 王明镜[1] 明静 王德好 牛继聪 刘为易 胡晓梅[1] LI Yu-Jin;MA Ju-Ning;WANG Zi-Qin;YANG Er-Peng;WANG Ming-Jing;MING Jing;WANG De-Hao;NIU Ji-Cong;LIU Wei-Yi;HU Xiao-Mei(Department of Hematology,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Graduate School,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]中国中医科学院西苑医院血液科,北京100091 [2]北京中医药大学研究生院,北京100029

出　　处：《中国实验血液学杂志》2022年第3期836-843,共8页Journal of Experimental Hematology

基　　金：国家自然科学基金(82174360);中国中医科学院西苑医院国家自然科学基金培育项目(XY20-10);中国中医科学院科技创新工程(CI2021A01702)。

摘　　要：目的:基于蛋白组学技术分析原发性血小板增多症(ET)特异性蛋白标志物,寻找与血小板活化相关特异性蛋白,并进行验证。方法:应用非标记定量蛋白组学(LFP)检测ET患者和健康者血清蛋白质谱,选取疾病组蛋白丰浓度较对照组差异倍数上调或下调1.3倍及两组蛋白丰浓度经t检验P<0.05的蛋白定义为差异蛋白。应用KEGG和GO分析软件对差异蛋白进行生物信息学分析。组间差异蛋白平均丰浓度的差异分析采用t检验,P<0.05为差异有统计学意义。选取差异蛋白,应用平行反应监测进行验证。结果:本研究发现差异蛋白共140个,其中72个蛋白表达上调,68个蛋白表达下调。KEGG富集显示差异蛋白表达与血小板活化通路相关,与血小板活化相关的差异蛋白是GPV、COL1A2、GP1bα、COL1A1和GPVI,其中GPV、GP1bα和GPVI表达上调,COL1A2和COL1A1表达下调。对COL1A1、GP1bα、GPVI和GPV进行PRM验证,其结果与LFP蛋白组学结果一致。结论:ET患者差异蛋白与血小板活化通路激活相关,GPV、GPVI、COL1A1和GP1bα等差异蛋白可作为ET血小板活化相关的新靶点。Objective: To analysis the specific protein markers of essential thrombocythemia(ET) based on proteomics technology, to explore and verify the differential protein related to platelet activation. Methods: Blood samples were obtained from ET patients and healthy people and a certain protein mass spectrometry was detected using label-free quantitative technology. The proteins relative abundance increased or down-regulated by 1.3 times in the disease group compared with the control group, and the protein abundance in the two groups t test P<0.05 were defined as differential proteins. Bioinformatics analysis of the differential proteins was performed using GO and KEGG. The difference in the average protein abundance between the two groups was analyzed by t test and P<0.05 was considered statistically significant. Differential proteins were selected for verification by parallel reaction monitoring(PRM) technology. Results: A total of 140 differential proteins were found, of which 72 were up-regulated and 68 were down-regulated. KEGG enrichment showed that the differential protein expression was related to the platelet activation pathway. The differential proteins related to platelet activation were GPV, COL1A2, GP1bα, COL1A1 and GPVI. Among them, the expressions of GPV, GP1 bα and GPVI were up-regulated, and the expressions of COL1A2 and COL1A1 were down-regulated. PRM verification of COL1A1, GP1bα, GPVI and GPV was consistent with LFP proteomics testing. Conclusion: Differential proteins in ET patients are related to platelet activation pathway activation.Differential proteins such as GPV,GPVI,COL1A1 and GP1 bα can be used as new targets related to ET platelet activation.

关 键 词：原发性血小板增多症 血小板活化 蛋白组学 生物学标记 血清 

分 类 号：R558.3[医药卫生—血液循环系统疾病]