| In the post/genomic era, with the accomplishment of the sequence mapping of human genome, the proteomics will play a tremendous role. Proteomics or functional proteomics represents the array of proteins in a biological compartment(cell,tissue,organ) at a particular time, under a particular set of conditions. Proteomics is the integrated study of protein properties on a large scale and will facilitate the elucidation of network mechanism of the development, progress of the disease.Spinal cord injury(SCI) is a serious neurotrauma and the mechanism of injury and repair of SCI is very complicated. The application of proteomics technology can provide a new approach to reveal the molecular mechanisms of SCI. The proteomics studies about SCI is not reported. Proteomics will be very useful for developing diagnostic predictors after spinal cord injury and for mapping changes in proteins after injury in order to identify new therapeutic targets. The objective of study is to establish a immobilized pH gradient-based two-dimensional electrophoretic (2-DE) technology to detect and separate of spinal cord and cerebro spinal fluild (CSF) protein. By comparing the spinal cord and CSF proteome between SCI patient and controls , to identify protein that play a role in the disease process and thus to study the pathogenesis of SCI. Firstly , a animal model of SCI in rabbits was established with a modified Allen's method. Secondly, the CSF sample were obtained from the patient with SCI. and the control group. Thirdly , the total proteins of spinal cord and CSF were separated with immobilized pH gradient-based two-dimen-sional electro-phoretic. After silver staining gel werr acquired by scanner and analyzed with PD-Quest software. 2-DE maps of spinal cord and CSF were established. Partial differentially expressed protein spots were incised from gel and digested by trypsin in gel. LCQ-DECA XPplus –ES-MS and NCBInr database searching by SEQUEST software were used for protein identification. Well-resolved and reproducible 2-DE map of spinal cord and CSF were obtained with the modified 2-DE technology. Average sports of gels were 413±32 and 367±29 respectively in the spinal cord of control group and injury group with average matching rate of 85% and 83%. 40 differentially expressed protein sports were detected ,among which 19 were up-regulated ,11 were down-regulated, 6 were uniquely present and 4 disappeared in the injury group. Average sports of gels were131±10 and 144±16 respectively in the CSF of control group and injury group. 25 differentially expressed protein sports were detected ,among which 6 were up-regulated ,7 were down-regulated, 7 were uniquely present and 5 disappeared in the injury group. 12 spots in gel of spinal cord and 10 spots in CSF were further analysised by LCQ-DECA XPplus –ES-MS and identificated 9 and 7 proteins respectively. In the spinal cord group, Neurofilament triplet M protein,Nerve growth fator and Calpain were up-regulated but α1-Acid-glycoprotein and Oxygen regulated protein 1 were down-regulated. There were two newly expressed proteins Endooligopeptidase A related protein and hypothetical 152.1kDa protein as well as two disappeared proteins Calpain inhibitor and Hypothetical protein cgi-10. In the CSF group, Insulin-like growth factor I precursor,PRR2-alpha-humanⅡ and MIST were up-regulated butα-2-HS glycoprotein and hypothetical protein FLJ20309 was down-regulated. There were one disappeared proteins transferrin precusor as well as one newly expressed proteins Myelin basic protein Ⅱ.In this study , the animal model of SCI on different degree of impact established with a new weight-drop device has a stable and repetitive characters for the research on spinal cord injury. Well-resolved and reproducible 2-DE technological. patterns of spinal cord and CSF were established .The 2-DE map of spinal cord and CSF of SCI were obtained and particular differential protein were characterized. The results provide fundamental information for the molecular mecha...
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