Proteomics Study Of Brain Tissue In Model Of Intracerebral Hemorrhage In Rat

Objectives: Intracerebral hemorrhage(ICH) is a clinical common disease in nervous system, accounting for 20% to 30% in acute cerebrovascular disease. Its mortality and disability rate is very high and it has been severely damaged health. But there is no effective treatments yet. Clinic random comparision experiments have proved that clearing hemorrhage in the head counldn′t improve patients prognosis,and 1/3 patients still get worse after operation.This phenomnon hints there is secondary lesion around hemorrhage besides hemorrhage direct damage.Gene Function is .executed by proteins ultimately and proteins directly reflect the complexity and variability of life phenomena. Proteomics,the studying of dynamic changes of protein composition including cells, tissues and body fluids , expressions and modifications , the interaction and contact between the proteins, reveals protein functions and the laws of cellular life activities. Comparative proteomics offers the possibility of studying molecular mechanism in the total analysising differences in protein expressions.Our study plans to use comparative proteomics to research brain tissue proteins in ICH 72 hour and normal controls which may discover potential proteins about the disease.It may be a way to study the pathological and physiological mechanisms of ICH.It maybe can provide evidences of exploring the disease-specific markers and intervention targets. Methods:1.We made models of rat ICH.Kill the rats at 72 hour and got the tissues of the lesion side.Got the corresponding part in controls.2.The tissue proteins were extracted with mixture of urea,CHAPS,DTT,IPG buffer were run IPG isoelectric focusing electrophoresis as the first dimension,and then run vertical SDS-PAGE as the second dimension.The gels were visualized by colloidal coomassive blue staining and analysised with ImageMaster 2D Elite software,The proteins of interest were in-gel digested and identified using MALDI-TOF/TOF tandem mass spectrometry.Results:We made models of ICH in rat successfully.Our data showed that the levels of 39 proteins in brain tissue were significantly altered in ICH group compared with controls. 15 up-regulated protein spots were found which were identified as Glutamine synthetase,Heat shock cognate 71 kDa protein,F-actin-capping protein subunit alpha-2,Apolipoprotein E,Triosephosphate isomerase,Dihydropyrimidinase-related protein2,NADH-ubiquinone oxidoreductase 75 Da subunit,Proteasome subunit alpha type-6,Glial fibrillary acidic protein,Cytochrome b-c 1complex subunit 1,Mu-crystallin homolog,Annexin A5,prohibitin,Alpha-soluble NSF attachment protein.While 24 down-regulated proteins were identified as Transgelin-3, ATP synthase subunit d, Phosphoglycerate mutase 1, Pyridoxal kinase, Aspartate aminotransferase, Tyrosine 3-monooxygenase, Alpha-enolase, Nucleoside diphosphate kinaseA, Nucleoside diphosphate kinase B, Superoxide dismutase(Cu-Zn), Glutathione S-transferase Yb-3, Cofilin-1, Fructose-bisphosphate aldolase C, Creatine kinase B-type, 4-aminobutyrate aminotransferase, Neurofilament light polypeptide, Aconitate hydratase, Peroxiredoxin-2 , COP9 signalosome complex subunit 4.Conclusion:1.We got 2-DE maps of the whole proteins from the brain tissues with ICH group and normal controls.2.The identified proteins including cytoskeleton proteins, energy metabolic enzymes,antioxidants proteins and apoptosis related proteins. We concluded that these proteins may be involved in the pathogenesis of ICH, but the role of these proteins still remains to be elucidated in detail and further validation is needed, and established fundament for explored the disease-specific markers and intervention targets of ICH.