Comparative Proteomic Study Of Epileptic Foci In Human Patients With Epilepsy

Objective:Epilepsy is a paroxysmal brain dysfunction syndrome caused by highly synchronized abnormal discharge of the neurons, and it is one of the common diseases of the nervous system, the incidence of it is about5‰-7‰. Many reasons can cause epilepsy. Epilepsy with unknown reasons was called idiopathic epilepsy, and others have known reasons were called symptomatic epilepsy. The mechanism of epilepsy is complicated, nowadays theory of ion channels, abnormal networks ect have received much more attention. By this experiment we hope we can farther clarify the mechanism of the disease.Protein is the main carrier of biological function. Proteomic technology is a study that is based on the proteome nature and to clarify the mechanism of the disease, the cell mode and the functional relationships at protein level. By quantitative analysis and comparative study of protein expressions under different states, we have studied the variations with the protein expression and the modified changes, which will lead us to find the diagnostic markers and the assessment indicators.We researched the brain tissues of epileptics and controls by comparative proteomics, and the2D-DIGE spectrums were established. These differently expressed proteins were analyzed and identified, which may provide clues for further elucidate the mechanism and the therapeutic targets of epilepsy.Methods:1. The brain tissue proteins were separated by2D-DIGE, then gels were scanned on the Typhoon multifunctional laser scanning imaging system and pictures were obtained by ImageMaster software, differently expressed protein spots were picked out.2. These proteins were analyzed by MALDI-TOF-MS technology and PMFs were acquired. Finally, these differential proteins were identified through protein databases with Mascot software.3. The differential expression levels of glial fibrillary acidic protein and cofilin-1were verified by western blot.Result:1.48differential expressed proteins were picked out and15were identified by the NCBI database,13were down-regulated and2were up-regulated in the patients.2. These proteins were classified according to their functions, which related to neuroprotection, cytoskeleton, oxidative stress, amino acid metabolism, ion channels, energy metabolism and so on. They may play an important role in the pathogenesis of epilepsy.3. glial fibrillary acidic protein was down-regulated and cofilin-1was up-regulated in epileptic patients.Conclusion:The2D-DIGE pictures of epileptics were established, and the differently expressed proteins were identified, which may be beneficial for further elucidated the mechanism of epilepsy.