Comparative Proteomic Study Of Cerebrospinal Fluid In Epilepsy Patients

Objective:The aim of this study is to establish the proteomic system of eerebrospinal fluid. Through studying the comparative proteomics of cerebrospinal fluid in generalized tonic-clonic seizure, status epilepticus and temporal lobe epilepsy related to hippoeampal sclerosis patients, to find the biomarkers of epilepsy patients, investigate the pathological mechanisms of the epilepsy, and search new drug targets.Method:The CSF were extracted from the patients of headache, the generalized tonic-clonic seizure, status epilepticus and temporal lobe epilepsy related to hippoeampal sclerosis. The CSF were enriched protein through ultrafiltration and removed albumin and IgG by removal kit. The concentration of protein was quantitated with Bradford method. The electrophoresis included immobilized pH gradients isoelectric focusing electrophoresis and sodium dodecyl sulfate polyacrylamide gels electrophoresis. The gels were stained with silver nitrate and Coomassico brilliant blue. The gels were analyzed by ImageMaster 2D Platinum 5.0 software. The differential proteins were identified in SW1SS-2DPAGE database and the interested proteins in-gels were digested and identified using MALDI-TOF-MS.Result:1. This study establish the proteomic technique system of cerebrospinal fluid.2. We found six up-regulated protein spots in 2-D gels of generalized tonic-clonic seizure patients compared with headache patients, which were identified as Haptoglobin and Zinc finger protein 605, while fourteen down-regulated protein spots in 2-D gels of GTCS patients, which were identifiedas Haptoglobin, Plasma retinol-binding protein, Zinc finger protein 613.3. We found eighteen up-regulated protein spots in 2-D gels of status epilepticus patients compared with generalized tonic-clonic seizure patients, which were identified as Leucine-rich alpha-2-glycoprotein, Clusterin (Complement-associated protein SP-40, 40) (Complement cytolysis inhibitor) (Apolipoprotein J) (Apo-J), Apolipoprotein A-I, Apolipoprotein D, Haptoglobin, Immunoglobulin light chain, Serum albumin precursor, tRNA pseudouridine synthase B, T-cell receptor beta chain V region CTL-L17 precursor, Gamma crystallin C.4. We found twenty-five up-regulated protein spots in 2-D gels of temporal lobe epilepsy related to hippocampal sclerosis compared with generalized tonic-clonic seizure patients, which were identified as Transthyretin (Prealbumin) (TBPA), Immunoglobulin heavy chain gamma, Complement C4-B, Deoxycytidylate deaminase. while five down-regulated protein spots in 2-D gels of TLE-HS patients, which were identified as Zinc finger protein 605, and Zinc finger CCHC domain-containing protein 5.Conclusion:Through studying the epilepsy patients CSF proteins by comparative proteomics, we got a number of related proteins of epilepsy. It is very useful for finding the biomarkers and new drug targets of epilepsy, and understanding the pathological mechanisms and pathophysiological process of the epilepsy.