Profiling the proteomic inflammatory state of human astrocytes using DIA mass spectrometry.
Profiling the proteomic inflammatory state of human astrocytes using DIA mass spectrometry.
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使用DIA质谱分析人类星形胶质细胞的蛋白质组学炎症状态。
DOI：
10.1186/s12974-018-1371-6
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发表时间：
2018-11-30
影响因子：
9.3
通讯作者：
Sanchez JC
中科院分区：
文献类型：
--
作者：
Dozio V;Sanchez JC
星形胶质细胞是中枢神经系统中最丰富的细胞，对正常的神经元发育、突触形成、血脑屏障调节和大脑稳态具有重要作用。它们还积极参与启动和维持神经炎症反应。然而，关于炎症下它们的蛋白质组表型的信息目前是有限的。数据独立获取质谱法应用于广泛表征在TNF， IL-1β和LPS诱导的不同炎症条件下永生化人胎儿星形胶质细胞中超过4000种蛋白质的特征，而基于多重免疫测定的筛选用于量化在这些炎症条件下释放的广泛细胞因子。然后采用免疫细胞化学和免疫印迹法验证不同刺激条件下典型和非典型NF-κB的活化情况。最后，我们建立了一个体外血脑屏障模型，该模型由原代人脑微血管内皮细胞和原代人星形胶质细胞共同培养组成，用于验证星形胶质细胞在更复杂的体外系统中暴露于LPS后的炎症反应。我们报道了一组186个蛋白，它们的水平被TNF、IL-1β和LPS显著调节。这三种刺激诱导蛋白质组紊乱，导致参与抗原呈递和非典型NF-κB通路的关键炎症蛋白丰度增加。TNF和IL-1β，而非LPS，也激活了典型的NF-κB通路，这反过来导致广泛的炎症反应和细胞骨架和粘附蛋白的失调。此外，TNF和LPS（而非IL-1β）增加了几种干扰素刺激基因产物的丰度。最后，TNF和IL-1β同样上调了几种细胞因子和趋化因子的分泌，而LPS仅诱导IL-8、IFN-γ和IL-1β分泌适度增加。在与暴露于LPS的脑微血管内皮细胞共培养的原代星形胶质细胞中，也观察到与I型IFN和非典型NF-κB信号相关的蛋白上调。本研究提供了关于人类星形胶质细胞暴露于炎症刺激下的蛋白质组学表型的综合信息，包括单培养和与人脑微血管内皮细胞共培养。本文的在线版本（10.1186/s12974-018-1371-6）包含补充材料，授权用户可使用。
Astrocytes are the most abundant cells in the central nervous system and are responsible for a wide range of functions critical to normal neuronal development, synapse formation, blood-brain barrier regulation, and brain homeostasis. They are also actively involved in initiating and perpetuating neuroinflammatory responses. However, information about their proteomic phenotypes under inflammation is currently limited. Data-independent acquisition mass spectrometry was applied to extensively characterize the profile of more than 4000 proteins in immortalized human fetal astrocytes under distinct inflammatory conditions induced by TNF, IL-1β, and LPS, while multiplex immunoassay-based screening was used to quantify a wide range of cytokines released under these inflammatory conditions. Then, immunocytochemistry and western blotting were used to verify the activation of canonical and non-canonical NF-κB upon exposure to the different stimuli. Finally, an in vitro model of the blood-brain barrier consisting of a co-culture of primary human brain microvascular endothelial cells and primary human astrocytes was used to verify the inflammatory response of astrocytes upon LPS exposure in a more complex in vitro system. We reported on a set of 186 proteins whose levels were significantly modulated by TNF, IL-1β, and LPS. These three stimuli induced proteome perturbations, which led to an increased abundance of key inflammatory proteins involved in antigen presentation and non-canonical NF-κB pathways. TNF and IL-1β, but not LPS, also activated the canonical NF-κB pathway, which in turn led to an extensive inflammatory response and dysregulation of cytoskeletal and adhesion proteins. In addition, TNF and LPS, but not IL-1β, increased the abundance of several interferon-stimulated gene products. Finally, TNF and IL-1β similarly upregulated the secretion of several cytokines and chemokines, whereas LPS only induced a moderate increase in IL-8, IFN-γ, and IL-1β secretion. Upregulation of proteins associated with type I IFN and non-canonical NF-κB signaling was also observed in primary astrocytes co-cultured with primary brain microvascular endothelial cells exposed to LPS. The present study provides comprehensive information about the proteomic phenotypes of human astrocytes upon exposure to inflammatory stimuli both in monoculture and in co-culture with human brain microvascular endothelial cells. The online version of this article (10.1186/s12974-018-1371-6) contains supplementary material, which is available to authorized users.
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影响因子：
3.7
作者：
Choi, Su-Lim;Lee, Sang Jun;Lee, Seung-Goo
通讯作者：
Lee, Seung-Goo
DOI：
10.1073/pnas.89.16.7840
发表时间：
1992-08-15
影响因子：
11.1
作者：
FU, XY;SCHINDLER, C;DARNELL, JE
通讯作者：
DARNELL, JE
影响因子：
11.4
作者：
Cheon, HyeonJoo;Holvey-Bates, Elise G.;Schoggins, John W.;Forster, Samuel;Hertzog, Paul;Imanaka, Naoko;Rice, Charles M.;Jackson, Mark W.;Junk, Damian J.;Stark, George R.
通讯作者：
Stark, George R.
DOI：
10.1016/j.biocel.2004.10.004
发表时间：
2005-06-01
影响因子：
4
作者：
Gee, JR;Keller, JN
通讯作者：
Keller, JN
影响因子：
82.9
作者：
Herold, Nikolas;Rudd, Sean G.;Schaller, Torsten
通讯作者：
Schaller, Torsten