Cardiac myocyte exosomes: stability, HSP60, and proteomics
Cardiac myocyte exosomes: stability, HSP60, and proteomics
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DOI：
10.1152/ajpheart.00835.2012
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发表时间：
2013-04-01
影响因子：
4.8
通讯作者：
Knowlton, A. A.
中科院分区：
文献类型：
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作者：
Malik, Z. A.;Kott, K. S.;Knowlton, A. A.
Malik ZA，Kott KS，Poe AJ，Kuo T，Chen L，费拉拉KW，诺尔顿AA.心肌细胞外泌体：稳定性，HSP 60和蛋白质组学。Am J Physiol Heart Circ Physiol 304：H954-H965，2013。首次发表于2013年2月1日; doi：10.1152/ajpheart.00835.2012.-外泌体是直径为50至100 nm的脂质囊泡，与细胞间通讯有关，包括传播恶性肿瘤，并作为病毒颗粒在传播到新细胞时逃避检测的一种方式。以前，我们证明了成年心肌细胞在外泌体中释放热休克蛋白（HSP）60。当不在外泌体中时，细胞外HSP 60通过激活Toll样受体4引起心肌细胞凋亡。因此，从外泌体释放HSP 60将损害周围的心肌细胞。我们假设1）环境中的病理变化，例如发热、pH变化或乙醇消耗，会增加外泌体渗透性; 2）不同的外泌体诱导剂会导致不同的外泌体蛋白含量; 3）“生理”浓度的乙醇会导致外泌体释放;以及4）ROS产生是外泌体产生增加的潜在机制。我们发现以下内容：首先，基于蛋白质印迹分析，外来体在不同的生理/病理条件下保留它们的蛋白质货物。第二，质谱分析表明，心脏外泌体的蛋白质含量与文献中其他类型的外泌体显著不同，并且含有胞质、肌节和线粒体蛋白。第三，乙醇不影响外泌体的稳定性，但大大增加了心肌细胞外泌体的产生。第四，乙醇和缺氧/复氧来源的外泌体具有不同的蛋白质含量。最后，ROS抑制减少了外泌体的产生，但并没有完全抑制它。总之，外泌体蛋白质含量的影响，细胞来源和刺激外泌体形成。ROS刺激外泌体产生。外泌体的功能仍有待充分阐明。
Malik ZA, Kott KS, Poe AJ, Kuo T, Chen L, Ferrara KW, Knowlton AA. Cardiac myocyte exosomes: stability, HSP60, and proteomics. Am J Physiol Heart Circ Physiol 304: H954-H965, 2013. First published February 1, 2013; doi:10.1152/ajpheart.00835.2012.-Exosomes, which are 50- to 100-nm-diameter lipid vesicles, have been implicated in intercellular communication, including transmitting malignancy, and as a way for viral particles to evade detection while spreading to new cells. Previously, we demonstrated that adult cardiac myocytes release heat shock protein (HSP) 60 in exosomes. Extracellular HSP60, when not in exosomes, causes cardiac myocyte apoptosis via the activation of Toll-like receptor 4. Thus, release of HSP60 from exosomes would be damaging to the surrounding cardiac myocytes. We hypothesized that 1) pathological changes in the environment, such as fever, change in pH, or ethanol consumption, would increase exosome permeability; 2) different exosome inducers would result in different exosomal protein content; 3) ethanol at "physiological" concentrations would cause exosome release; and 4) ROS production is an underlying mechanism of increased exosome production. We found the following: first, exosomes retained their protein cargo under different physiological/pathological conditions, based on Western blot analyses. Second, mass spectrometry demonstrated that the protein content of cardiac exosomes differed significantly from other types of exosomes in the literature and contained cytosolic, sarcomeric, and mitochondrial proteins. Third, ethanol did not affect exosome stability but greatly increased the production of exosomes by cardiac myocytes. Fourth, ethanol- and hypoxia/reoxygenation-derived exosomes had different protein content. Finally, ROS inhibition reduced exosome production but did not completely inhibit it. In conclusion, exosomal protein content is influenced by the cell source and stimulus for exosome formation. ROS stimulate exosome production. The functions of exosomes remain to be fully elucidated.