Spray Stine, Johansson Sara E, Edwards Alistair V G, Larsen Martin R, Radziwon-Balicka Aneta, Povlsen Gro K, Edvinsson Lars
Department of Clinical Experimental Research, Glostrup Research Institute, Rigshospitalet, Glostrup, Denmark.
Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
J Mol Neurosci. 2017 Mar;61(3):396-411. doi: 10.1007/s12031-016-0875-8. Epub 2016 Dec 8.
This study aimed at obtaining an in-depth mapping of expressional changes of the cerebral microvasculature after transient global cerebral ischemia (GCI) and the impact on these GCI-induced expressional changes of post-GCI treatment with a mitogen-activated protein kinase kinase (MEK1/2) inhibitor. GCI was induced in male Wistar rats followed by treatment with either vehicle or the MEK1/2 inhibitor U0126 every 12 h post-GCI. Seventy-two hours after GCI or sham surgery, the cerebral microvasculature was isolated and the protein content analysed with state-of-the-art mass spectrometry. The proteomic profile of the isolated cerebral microvasculature 72 h after GCI (compared to sham) indicated that the main expressional changes could be divided into nine categories: (1) cellular respiration, (2) remodelling of the extracellular matrix, (3) decreased contractile phenotype, (4) clathrin-mediated endocytosis, (5) ribosomal activity, (6) expression of chromatin structure-related proteins, (7) altered synaptic activity, (8) altered G-protein signalling and (9) instability of the membrane potential. Treatment with U0126 partly normalized the expression of one or more of the proteins in all nine categories. Flow cytometry confirmed key findings from the proteome such as upregulation of the extracellular proteins lamininβ2 and nidogen2 (p < 0.05) after GCI. These results provide valuable molecular insight into the broad and complex expressional changes in the cerebral microvasculature after GCI and the effect of early MEK1/2 inhibitor treatment on these changes.
本研究旨在深入描绘短暂性全脑缺血（GCI）后脑微血管表达变化的图谱，以及丝裂原活化蛋白激酶激酶（MEK1/2）抑制剂对GCI诱导的这些表达变化的影响。在雄性Wistar大鼠中诱导GCI，然后在GCI后每12小时用赋形剂或MEK1/2抑制剂U0126进行治疗。GCI或假手术后72小时，分离脑微血管并使用最先进的质谱分析蛋白质含量。GCI后72小时（与假手术组相比）分离的脑微血管的蛋白质组学图谱表明，主要的表达变化可分为九类：（1）细胞呼吸，（2）细胞外基质重塑，（3）收缩表型降低，（4）网格蛋白介导的内吞作用，（5）核糖体活性，（6）染色质结构相关蛋白的表达，（7）突触活性改变，（8）G蛋白信号改变，（9）膜电位不稳定。用U0126治疗部分使所有九类中的一种或多种蛋白质的表达正常化。流式细胞术证实了蛋白质组的关键发现，例如GCI后细胞外蛋白层粘连蛋白β2和巢蛋白2的上调（p <0.05）。这些结果为GCI后脑微血管广泛而复杂的表达变化以及早期MEK1/2抑制剂治疗对这些变化的影响提供了有价值的分子见解。
