Proteomic Analysis Of Differential Expression Proteins Of Injured Spinal Cord Caused By Complete Transection

Objective: Spinal Cord Injury (SCI) is a severe neurotrauma. It could becaused by kinds of accidents, athletics and violence. After SCI, the feeling andmovement function will loss below the damage plane. How to improve theneuron regeneration and functional recovery are always a big problem in theworld. Consequently, the fact the factors which play essential roles in therecovery of SCI could not be specifically and effectively regulated alsocontribute to the limitations of the therapeutical application. However, as themechanism study of SCI proceeds, many evidences illustrate that CNS enjoysthe ability to regenerate. Many proteins associated with the improvement orinhibition of axon regeneration after SCI have gradually discovered, such asNeuron growth factor (NGF),neurotrophic factor-3 (NT-3),brain-derivedneurotrophic factor (BDNF) and Nogo. However, there are still many molecules participating in this progress have not been discovered. The purposeof our experiment is to find more important molecules involved in SCI usingproteomics technologies, and to illuminate the mechanisms involved in theprogress of injury and recovery in central nervous system.Methods: First, establish the complete spinal cord transaction modelswith SD rats. Then, the area of injured and normal spinal cords were dislodged,the Hydrophilicity and hydrophobicity proteins Were extracted. Theconcentrations of the proteins were detected by Bradford method. Then theproteins were separated by two-dimensional electrophoresis (2-DE). Thedifferentially expressed proteins were detected by PDQuest 7.1.1 software.After that, the interesting proteins were digested into peptides which could beanalyzed by matrix-assisted laser desorption -ionization quadrupoletime-of-flight tandem mass spectrometry (MALDI-Q-TOF MS/MS). We couldget the detailed protein information by searching the MS data throughMASCOT software. At last, the identified proteins were verified by RT-PCRand Western-blot.Results: The differentially expressed proteins were identified bymatrix-assisted laser desorption/ionization quadrupole time-of-flight massspectrometry (MALDI-Q-TOF MS), and a total of 18 differential expressionproteins were successfully identified. Three days after spinal cord injury, therewere 13 up-regulated proteins such as macrophage migration inhibitory factor(MIF),heat-shock protein 27 (HSP27),syndecan-3 (SDC3),T-cell receptorbeta chain precursorâ…¤region (CRTB4),annexinâ…¢(ANXâ…¢),galectin-3(LEG3),pyruvate dehydrogenase (ODPA),chromogranin A precursor(CgA),HSP70-binding protein 1 (HSPBP1). Meanwhile, triose-phosphate isomerase (TPIS), sphingosine 1-phosphate receptor 1 (S1P1),heat-shockprotein 10 (HSP10),peptidyl-prolyl cis-trans isomerase A (PPIase A) weredown-regulated. Most of the differentially expressed proteins have beenreported to participate in neuron growth, apoptosis and stress-reaction. Thedata will be helpful to illuminate the mechanisms involved in the progress ofinjury and recovery in central nervous system.