内质网
细胞生物学
免疫金标记
流式细胞术
细胞外
生物
绿色荧光蛋白
分泌物
细胞内
纳米粒子跟踪分析
污渍
化学
微泡
分子生物学
生物化学
解剖
超微结构
基因
小RNA
作者
Anna Koncz,Lilla Turiák,Krisztína Németh,Dorina Lenzinger,Tünde Bárkai,Péter Lörincz,Helga Zelenyánszki,Krisztina V. Vukman,Edit I. Buzás,Tamás Visnovitz
出处
期刊:Membranes
[MDPI AG]
日期:2023-04-13
卷期号:13 (4): 431-431
被引量:4
标识
DOI:10.3390/membranes13040431
摘要
Cardiomyopathies are leading causes of human mortality. Recent data indicate that the cardiomyocyte-derived extracellular vesicles (EVs) released upon cardiac injury are present in circulation. This paper aimed to analyze EVs released under normal and hypoxic conditions by H9c2 (rat), AC16 (human) and HL1 (mouse) cardiac cell lines. Small (sEVs), medium (mEVs) and large EVs (lEVs) were separated from a conditioned medium by a combination of gravity filtration, differential centrifugation and tangential flow filtration. The EVs were characterized by microBCA, SPV lipid assay, nanoparticle tracking analysis, transmission and immunogold electron microscopy, flow cytometry and Western blotting. Proteomic profiles of the EVs were determined. Surprisingly, an endoplasmic reticulum chaperone, endoplasmin (ENPL, grp94 or gp96), was identified in the EV samples, and its association with EVs was validated. The secretion and uptake of ENPL was followed by confocal microscopy using GFP-ENPL fusion protein expressing HL1 cells. We identified ENPL as an internal cargo of cardiomyocyte-derived mEVs and sEVs. Based on our proteomic analysis, its presence in EVs was linked to hypoxia in HL1 and H9c2 cells, and we hypothesize that EV-associated ENPL may have a cardioprotective role by reducing cardiomyocyte ER stress.
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