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模拟微重力诱导条件下大鼠下丘脑特定细胞防御蛋白的差异表达:比较蛋白质组学。

Differential expression of specific cellular defense proteins in rat hypothalamus under simulated microgravity induced conditions: comparative proteomics.

机构信息

School of Life Sciences, Beijing Institute of Technology, Beijing, P. R. China.

出版信息

Proteomics. 2014 Jun;14(11):1424-33. doi: 10.1002/pmic.201400019. Epub 2014 Apr 29.

DOI:10.1002/pmic.201400019
PMID:24648329
Abstract

Microgravity severely halts the structural and functional cerebral capacity of astronauts especially affecting their brains due to the stress produced by cephalic fluid shift. We employed a rat tail suspension model to substantiate simulated microgravity (SM) in brain. In this study, comparative mass spectrometry was applied in order to demonstrate the differential expression of 17 specific cellular defense proteins. Gamma-enolase, peptidyl-prolyl cis-trans isomerase A, glial fibrillary acidic protein, heat shock protein HSP 90-alpha, 10 kDa heat shock protein, mitochondrial, heat shock cognate 71 kDa protein, superoxide dismutase 1 and dihydropyrimidinase-related protein 2 were found to be upregulated by HPLC/ESI-TOF. Furthermore, five differentially expressed proteins including 60 kDa heat shock protein, mitochondrial, heat shock protein HSP 90-beta, peroxiredoxin-2, stress-induced-phosphoprotein, and UCHL-1 were found to be upregulated by HPLC/ESI-Q-TOF MS. In addition, downregulated proteins include cytochrome C, superoxide dismutase 2, somatic, and excitatory amino acid transporter 1 and protein DJ-1. Validity of MS results was successfully performed by Western blot analysis of DJ-1 protein. This study will not only help to understand the neurochemical responses produced under microgravity but also will give future direction to cure the proteomic losses and their after effects in astronauts.

摘要

微重力严重阻碍了宇航员的大脑结构和功能,特别是由于头部液流转移产生的压力,影响了他们的大脑。我们使用大鼠尾部悬吊模型来模拟大脑中的微重力。在这项研究中,比较质谱学被应用于证明 17 种特定细胞防御蛋白的差异表达。γ-烯醇酶、肽基脯氨酰顺反异构酶 A、神经胶质纤维酸性蛋白、热休克蛋白 HSP 90-α、10kDa 热休克蛋白、线粒体、热休克同源 71kDa 蛋白、超氧化物歧化酶 1 和二氢嘧啶酶相关蛋白 2 通过 HPLC/ESI-TOF 被发现上调。此外,通过 HPLC/ESI-Q-TOF MS 发现了 5 种差异表达的蛋白质,包括 60kDa 热休克蛋白、线粒体、热休克蛋白 HSP 90-β、过氧化物还原酶-2、应激诱导磷酸化蛋白和 UCHL-1。此外,下调的蛋白质包括细胞色素 C、超氧化物歧化酶 2、体细胞、兴奋性氨基酸转运体 1 和蛋白 DJ-1。通过 DJ-1 蛋白的 Western blot 分析成功验证了 MS 结果的有效性。这项研究不仅有助于了解微重力下产生的神经化学反应,还为治疗宇航员的蛋白质组损失及其后续影响提供了未来的方向。

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