大鼠脂肪间充质干细胞及其外泌体的蛋白组学对比分析  

Comparative proteomic analysis of rat adipose-derived mesenchymal stem cells and their exosomes

作　　者：张鑫瑞 韩悦 雷蕾 刘建宇 耿承奎[2] Zhang Xinrui;Han Yue;Lei Lei;Liu Jianyu;Geng Chengkui(Second School of Clinical Medicine,Yunnan University of Chinese Medicine,Chenggong Campus of Yunnan University of Chinese Medicine,Kunming 650000,Yunnan Province,China;2Department of Orthopedics,Yan’an Hospital Affiliated to Kunming Medical University,Kunming 650000,Yunnan Province,China)

机构地区：[1]云南中医药大学第二临床医学院,云南中医药大学呈贡校区,云南省昆明市650000 [2]昆明医科大学附属延安医院骨科,云南省昆明市650000

出　　处：《中国组织工程研究》2025年第13期2683-2689,共7页Chinese Journal of Tissue Engineering Research

基　　金：云南省科技厅科技计划项目(202001AY070001-263),项目负责人:耿承奎。

摘　　要：背景:从蛋白组学方面分析脂肪间充质干细胞及其来源外泌体促进组织修复的研究较多,但缺少二者之间的蛋白组学对比分析。目的:对脂肪间充质干细胞及其外泌体进行蛋白组学分析。方法:分离培养大鼠脂肪间充质干细胞,从细胞上清液中提取外泌体并进行鉴定;运用DIA蛋白组学分析大鼠脂肪间充质干细胞及其外泌体差异表达蛋白,对差异表达蛋白进行GO和KEGG分析。结果与结论:①脂肪间充质干细胞以长梭形为主,类似成纤维样;②经BCA法测定外泌体悬液的蛋白质量浓度为7.66 mg/mL;③外泌体呈典型的茶杯型,可见双层膜的囊泡结构,中央为低电子密度成分,外泌体粒径分布峰值为112.2 nm,浓度为7.5×10^(11)粒子/mL,平均直径在70-140 nm之间;④外泌体有表面标记蛋白CD9、TSG101高表达;⑤GO分析脂肪间充质干细胞及其外泌体的差异表达蛋白主要集中在细胞外基质和突触等位置,与离子结合和核糖体的结合能力相关,在细胞黏附和翻译过程中最为显著;⑥KEGG通路分析结果显示,差异表达蛋白主要参与细胞外基质受体交互、核糖体和细胞因子受体相互作用等,并且与胆固醇和甲状腺等多种代谢性疾病有关。BACKGROUND:Research on adipose-derived mesenchymal stem cells and their exosomes often uses proteomics to analyze their roles in tissue repair,but comparative proteomic analyses between the two are scarce.OBJECTIVE:To analyze adipose-derived mesenchymal stem cells and their exosomes using proteomic analysis.METHODS:Rat adipose-derived mesenchymal stem cells were isolated,cultured,and identified.Exosomes were then extracted from cell supernatant and identified.Differentially expressed proteins of adipose-derived mesenchymal stem cells and their exosomes were analyzed using DIA proteomics.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed on these differentially expressed proteins.RESULTS AND CONCLUSION:(1)Adipose-derived mesenchymal stem cells displayed a mainly spindle-shaped,fibroblast-like morphology.(2)The exosome suspension protein concentration was 7.66 mg/mL,as determined by the BCA method.(3)Exosomes exhibited a characteristic teacup shape with a visible double-layer membrane vesicle structure.The center presented a low electron density component.The exosomes showed a peak particle size distribution of 112.2 nm,a concentration of 7.5×10^(11) particles/mL,and an average diameter ranging from 70 to 140 nm.(4)Exosomes expressed high levels of surface marker proteins CD9 and TSG101.(5)Gene Ontology analysis of differentially expressed proteins revealed enrichment in the extracellular matrix and synapses,with functions related to ion binding,ribosome binding,and particularly cell adhesion and translation.(6)Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that the differentially expressed proteins were primarily involved in extracellular matrix receptor interaction,ribosome,and cytokine receptor interaction,and also associated with various metabolic diseases like cholesterol and thyroid disorders.

关 键 词：脂肪间充质干细胞 外泌体 蛋白组学 对比分析 组织修复 

分 类 号：R459.9[医药卫生—治疗学] R-332[医药卫生—临床医学] R318.08