中国输血杂志2025，Vol.38Issue(10)：1299-1306,8.DOI:10.13303/j.cjbt.issn.1004-549x.2025.10.001
单采血小板中囊泡亚型的蛋白质组学特征与功能调控研究
Proteomic characteristics and functional regulation of vesicle subtypes in apheresis platelets
摘要
Abstract
Objective To detect the different proteomic characteristics of microvesicles(MVs)and exosomes(EXOs)released from apheresis platelets during storage,and to explore their role in mediating platelet storage damage lesion(PSL).Methods Apheresis platelets were collected from the retention bag on the third day of storage.MVs and EXOs were isola-ted using differential centrifugation.Platelet,MVs and EXOs protein samples were extracted respectively,and the differen-tially expressed proteins were detected by quantitative proteomics technology.Further,the co-incubation model of MVs,EXOs and fresh platelets was adopted to evaluate the effect of extracellular vesicles on PSL.The aggregation response of platelets to collagen agonizers and the changes in ATP release rate were evaluated by optical turbidimetry.Flow cytometry was used to evaluate the changes of platelet early activation indicators(P-selectin and PAC-1)and mitochondrial membrane potentia.Western blot was used to detect the changes in the expression of key proteins for platelet activation and apoptosis(P-selectin,Integrin β3 and Bcl-xl).Results Proteomic analysis revealed a significantly separation in protein expression profiles of platelet,MVs and EXOs samples within the latent variable space.Energy metabolization-related proteins such as mitochondrial respiratory chain complex and oxidative phosphorylation were enriched specifically,in MVs while EXOs were enriched with inflammation-related proteins.Co-incubation experiments confirmed that extracellular vesicles could signifi-cantly induce platelet responses to agonists(the maximum aggregation rate in the MVs group increased by 187.36%,P<0.001;71.26%,in the EXOs group P=0.002).The maximum ATP release rate of platelets also increased(275.44%in the MVs group,P<0.001;70.18%in the EXOs group,P=0.015).The expression of P-selectin increased(119.33%in the MVs group,P<0.001;25.61%in the EXOs group,P=0.013),as detected by flow cytometry.The binding rate of PAC-1 increased(132.18%in MVs group,P<0.001;21.41%in EXOs group,P=0.043),and the mitochondrial mem-brane potential decreased(20.49%in MVs group,P<0.001;9.73%in EXOs group,P=0.044).In the MVs group,platelet P-selectin and Integrin β3 expression were significantly increased(100.83%and 395.64%,P<0.001),while Bcl-xl expression was lower than that in the control group(83.94%,P<0.001).Compared with the control group,P-se-lectin and Integrinβ3 expression were also increased(27.89%and 181.91%,P=0.007 和P=0.002),while Bcl-xl was decreased in the EXOs group(36.52%,P<0.001).Conclusion MVs and EXOs derived from stored platelets show dif-ferent proteomic characteristics.Compared with EXOs,MVs exhibits a stronger effect in inducing mitochondrial dysfunc-tion.Mvs also promots PSL responses including platelet activation and apoptosis.关键词
单采血小板/外泌体/微囊泡/储存损伤/线粒体膜电位Key words
apheresis platelets/exosomes/microvesicles/storage lesion/mitochondrial membrane potential分类
医药卫生引用本文复制引用
程红,胡作建,王佳琦,李丹丹,王志成,夏荣..单采血小板中囊泡亚型的蛋白质组学特征与功能调控研究[J].中国输血杂志,2025,38(10):1299-1306,8.基金项目
国家自然科学基金面上项目(82470234) （82470234）
中国输血协会圣湘基金(CSBT-SX-2024-05) （CSBT-SX-2024-05）
