A gel-based proteomic comparison of human cerebrospinal fluid between inflicted and non-inflicted pediatric traumatic brain injury
A gel-based proteomic comparison of human cerebrospinal fluid between inflicted and non-inflicted pediatric traumatic brain injury
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DOI：
10.1089/neu.2006.0061
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发表时间：
2007-01-01
影响因子：
4.2
通讯作者：
Kochanek, Patrick M.
中科院分区：
文献类型：
--
作者：
Gao, Wei-Min;Chadha, Mandeep S.;Kochanek, Patrick M.
创伤性脑损伤是婴幼儿最常见的创伤性死亡原因，而致命性脑损伤是导致婴幼儿外伤性死亡的主要原因。与其他中枢神经系统病理一样，脑损伤改变了脑脊液(CSF)的成分，这可能是研究脑病理生理的一个独特的临床窗口。采用双向凝胶电泳法(DIGE)结合质谱仪(MS)对13例重型颅脑损伤(n=13)和13例非重型颅脑损伤(n=13)患儿的脑脊液蛋白质谱进行了比较。用不同的荧光染料对iTBI和nTBI的脑脊液蛋白进行三联荧光标记，并用双向凝胶电泳法进行分离。在脑脊液中发现了大约250个蛋白质点，凝胶间2-D凝胶重复性达到90%。凝胶内消化后，用MS分析胰蛋白酶多肽以进行蛋白质鉴定。急性期的反应物，结合珠蛋白(HP)亚型，显示出nTBI比iTBI增加了大约四倍。相比之下，脑损伤后前列腺素D-2合成酶(PGDS)和半胱氨酸氨基转移酶(CC)的水平分别是非脑损伤的12倍和7倍。免疫印迹法检测Hp、PGDS、CC的变化。这些基于传统凝胶的蛋白质组学的初步结果显示了新的蛋白质变化，最终可能有助于理解iTBI和nTBI之间的病理生理学差异。
Traumatic brain injury (TBI) is the most common cause of traumatic death in infancy, and inflicted TBI (iTBI) is the predominant cause. Like other central nervous system pathologies, TBI changes the composition of cerebrospinal fluid (CSF), which may represent a unique clinical window on brain pathophysiology. Proteomic analysis, including two-dimensional (2-D) difference in gel electrophoresis (DIGE) combined with mass spectrometry (MS), was used to compare the CSF protein profile of two pooled samples from pediatric iTBI (n = 13) and non-inflicted TBI (nTBI; n = 13) patients with severe injury. CSF proteins from iTBI and nTBI were fluorescently labeled in triplicate using different fluorescent Cy dyes and separated by 2-D gel electrophoresis. Approximately 250 protein spots were found in CSF, with 90% between-gel reproducibility of the 2-D gel. Following in-gel digestion, the tryptic peptides were analyzed by MS for protein identification. The acute phase reactant, haptoglobin (HP) isoforms, showed an approximate fourfold increase in nTBI versus iTBI. In contrast, the levels of prostaglandin D-2 synthase (PGDS) and cystatin C (CC) were 12-fold and sevenfold higher in iTBI versus nTBI, respectively. The changes of HP, PGDS, and CC were confirmed by Western blot. These initial results with conventional gel-based proteomics show new protein changes that may ultimately help to understand pathophysiological differences between iTBI and nTBI.