青年与中年男性血浆外泌体蛋白组学差异浅析*

doi:10.3969/j.issn.1671-2587.2024.03.004

临床输血与检验 ›› 2024, Vol. 26 ›› Issue (3): 315-325.DOI: 10.3969/j.issn.1671-2587.2024.03.004

青年与中年男性血浆外泌体蛋白组学差异浅析^*

肖潘¹, 刘瑜琳², 薛晓楠¹, 薛英娜¹, 刘羿¹, 孙莉萍¹, 于洋¹

¹中国人民解放军总医院第一医学中心输血医学科,北京 100853;
²广安市人民医院输血科,四川广安 638500

收稿日期:2024-03-24 出版日期:2024-06-20 发布日期:2024-06-24
通讯作者: 于洋,主要从事单采治疗、野战输血及智能化输血辅助策略系统研究,（E-mail）yuyangpla301@163.com。共同通信作者：孙莉萍,主要从事输血医学基础研究,（E-mail）sunlp2012@sina.com。
作者简介:肖潘,主要从事输血医学研究,（E-mail）sirius_xp@sina.cn。
基金资助:
*国家自然科学基金项目（No. 81970167）资助

Proteomic Analysis of Plasma Exosomes from Young Versus Middle-aged People

XIAO Pan¹, LIU Yulin², XUE Xiaonan¹, XUE Yingna¹, LIU Yi¹, SUN Liping¹, YU Yang¹

¹Department of Transfusion Medicine, The First Medical Center, Chinese PLA General Hospital, Beijing 100853;
²Transfusion Department, Guang'an People's Hospital, Guangan 638500

Received:2024-03-24 Online:2024-06-20 Published:2024-06-24

摘要/Abstract

摘要： 目的外泌体作为细胞信使,在体液循环中运输各种生物活性分子。本研究初步对比青年和中年男性血浆外泌体的性状特征和内含蛋白质的差异,为探索血浆外泌体内对机体有影响的蛋白质提供一定的理论指导。方法收集青年（19.33岁±1.16岁）与中年（50.33岁±2.52岁）男性的外周血,通过差速超速离心法分离血浆外泌体,利用透射电镜、粒径分析仪和Western blot对其表征进行检测。液相质谱分析技术检测血浆外泌体蛋白,对比分析两个年龄层来源的血浆外泌体蛋白种类与功能差异。结果青年男性血浆外泌体的平均直径和蛋白质浓度与中年男性的血浆外泌体相近,但外泌体的平均浓度略低于中年男性;青年男性血浆外泌体低表达CD9,而中年男性则呈现高表达。在两组人群的血浆外泌体中共鉴定到110个差异蛋白;相较于中年男性,青年男性血浆外泌体有36个蛋白表达上调,74个蛋白表达下调。GO功能分析显示差异蛋白的生物学过程差异主要集中在翻译延伸、细胞大分子生物合成和有机氮化合物生物合成;分子功能主要集中在翻译延伸因子、铜离子结合和核酸结合。KEGG通路分析显示差异蛋白主要富集在抗原处理、呈递和内吞作用等13条通路。在仅青年男性血浆外泌体表达的21个蛋白中,Drebrin-like protein（DBNL）同时具有神经系统和免疫系统的相关功能。结论尽管青年和中年男性血浆外泌体在形态、粒径和浓度上无明显差异,但其表面标志物和内含蛋白质却存在显著差异。

关键词: 血浆, 外泌体, 蛋白组学, 年龄

Abstract: Objective Exosomes, as cellular messengers circulating in bodily fluid, carry diverse bioactive chemicals. This study aimed to analyze the characteristics and variations in plasma exosomal proteins between young and middle-aged individuals, offering theoretical insights into their role in influencing bodily functions. Methods Peripheral blood samples were collected from young (age:19.33±1.16) and middle-aged (age:50.33±2.52) adults. Plasma exosomes were isolated by differential ultracentrifugation, and characterized via transmission electron microscopy, particle size analysis, and Western blot. Liquid-phase mass spectrometry analysis technology was used to detect and analyze the types and functional differences of plasma exosome proteins between the two age groups. Results The average diameter and protein concentration of plasma exosomes in young men were similar to those in middle-aged men, but the average concentration of exosomes was slightly lower in young men. Plasma exosomes from young men exhibited low CD9 levels, contrary to high CD9 levels observed in middle-aged men. A total of 110 differential proteins were identified in the plasma exosomes of both groups, with 36 proteins upregulated and 74 downregulated in young males compared to middle-aged males. GO functional analysis revealed differences in biological processes, particularly translation elongation, cellular macromolecule biosynthesis, and organic nitrogen compound biosynthesis. Molecular functions focused on translation elongation factors, copper ion binding, and nucleic acid binding. KEGG pathway analysis indicated enrichment in 13 pathways, including antigen processing, presentation, and endocytosis. Among the 21 proteins specific to young men's plasma exosomes, Drebrin-like protein (DBNL) displayed functions related to both the nervous system and the immune system. Conclusion No differences have been noted in the morphology, size, and concentration of plasma exosomes between young and middle-aged men, but there are significant differences in their surface markers and intronic proteins.

Key words: Plasma, Exosome, Proteomics, Age

中图分类号:

R457

肖潘, 刘瑜琳, 薛晓楠, 薛英娜, 刘羿, 孙莉萍, 于洋. 青年与中年男性血浆外泌体蛋白组学差异浅析^*[J]. 临床输血与检验, 2024, 26(3): 315-325.

XIAO Pan, LIU Yulin, XUE Xiaonan, XUE Yingna, LIU Yi, SUN Liping, YU Yang. Proteomic Analysis of Plasma Exosomes from Young Versus Middle-aged People[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2024, 26(3): 315-325.

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青年与中年男性血浆外泌体蛋白组学差异浅析^*

Proteomic Analysis of Plasma Exosomes from Young Versus Middle-aged People

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