摘要
目的本研究旨在探查临床可介入性生物样本脑脊液中锰毒性相关的差异表达蛋白谱,为今后深入的毒性机制研究和病程相关生物标志研究提供科学数据。方法运用非标记(label free)定量蛋白质组学技术分析比较成年大鼠经腹腔注射暴露于氯化锰(以锰计,6 mg Mn/kg BW)1个月(亚急性)、3个月(亚慢性)及3个月加30 d恢复期后与其相应对照组(腹腔注射生理盐水)大鼠的脑脊液中蛋白质的差异表达情况,并根据蛋白质的基因本体(gene ontology,GO)注释对所鉴定蛋白进行了分类分析。结果本研究共鉴定到173个蛋白质,经差异性比较与重复蛋白剔除后获得123个锰毒性相关的差异表达蛋白,其中,染锰后55个蛋白质表达量上调,68个蛋白质表达量下调。根据差异性,123个差异表达蛋白可分为4个层次,分别为表达升高(4个),表达下降(2个),仅在对照组鉴定到(66个),仅在染毒组鉴定到(51个)。对123个差异蛋白按其GO注释分别进行细胞组件、分子功能及生物学过程分类后发现,差异表达蛋白主要来自于细胞核、细胞外基质及细胞质,以结合功能为主,少数参与代谢、应对刺激等生物学过程,半数以上的生物学过程未知。结论由于脑脊液中的蛋白质多数都是由脉络丛组织合成分泌的,因此上述脑脊液中差异表达的蛋白质组不仅可以作为锰神经毒性作用引起的神经退行性疾病的潜在的生物学标志,而且还为锰致脉络丛(血-脑脊液屏障的组织基础)上皮细胞毒性损伤作用分子机制的深入研究提供线索和实验基础。
Objective The regulation of brain Mn depends largely on the blood-brain barrier and blood-cerebrospinal fluid barrier （BCB）. The latter is constituted by choroid plexus （CP） epithelial cells, which is specialized for cerebrospinal fluid （CSF） production, has been considered as a primary target in Mn-induced neurotoxicity. This study aims to explore the differential proteome in CSF after sub-acuteand sub-chronic Mn exposure in SD rats. Methods （ 1 ） Establishment of Mn-animal Model： Rats （ 1.5 month） were divided into 6 groups ; each received daily ip injections of either MnC12 （6 mg Mn/kg BW） or saline （ as controls） for 30days （ sub-acute）, 90 days （ sub- chronic）, or 90 days followed by additional 30-day convalescence. The CSF were collected and analyzed. （2） Evaluation of Mn-animal model （ data not shown here, which will be published in another paper）. （ 3 ） Mn-related proteome in CSF was analyzed by a novel proteomic technique of label-free nanoHPLC-Q-TOF-MS/MS. Results A total of 123 Mn-related differential proteins in CSF were identified, of which 55 were up-regulated, 68 were down-regulated. These identified differential proteins could be divided into three categories according to their characteristic change fold （Mn vs. Control）： （1） 4 proteins＇ expression increased by change fold 〉 1.5, （2） 2 porteins＇ expression decreased by change fold 〉 1.5, （3） 66 proteins were only detected in the control group, （4） 51 proteins were only detected in the Mn groups. Based on the information of GO categories, these differentially expressed proteins were mainly from the nuclei, involving in the function of binding; however, more than half of the proteins have no characterized biological function. Conclusion Considering of majority of proteins in CSF being produced and excreted by CP, these differential proteins may be valuable for exploring the novel potential biomarkers to diagnose and monitor the progression of Mn-indueed neurodegenerative disease clinically ; meanwhile, the results shed light on the future molecular mechanism study of Mn on choroids plexus epithelial cells.
出处
《毒理学杂志》
CAS
CSCD
北大核心
2012年第4期247-253,共7页
Journal of Toxicology
基金
北京市留学人员科技活动择优资助项目(2007-62)
2007市属公益院所改革与发展项目(2007)
北京市"十百千"卫生人才培养专项(2007年"百"层次
2009年"十"层次)
北京市卫生系统高层次卫生技术人才培养项目(2011)
国家重大科技专项项目子课题(2008ZX10002-106)
