Rearch Of Differentially Expressed Protein In The Brain Of Model Insomnia Rats By Acupuncture Based On Mass-spectrometric Technique

Objective：Use proteomics methods through bio-mass spectrometry techniques toidentify differences in protein molecular weight and protein content, Swiss-Portprotein database to retrieve, finding and screening associated with insomnia andacupuncture treatment of insomnia associated differentially expressed proteins, fromthe perspective of proteomics to explore insomnia pathogenesis and mechanism ofaction of acupuncture treatment of insomnia and provide experimental basis.Methods：1. Selection of clean grade male SD rats150were randomly dividedinto five groups: control group, normal bundle control group,normal acupuncturegroup, model control group,model acupuncture group.Modeling method usingintraperitoneal injection Chlorophenylalanine suspension.2.After the end of modeling the rats were decapitated, brains tissue wereremoved, five groups of rats were extracted from whole brain total protein, proteinconcentration was determined; using two-dimensional electrophoresis separation andpurification of the brain protein, followed by the Coomassie brilliant blue staining,then scanning the gel image to obtain the two-dimensional gel electrophoresis proteinatlas. application of2-DE map analysis software "ImageMaster2D Platinum" ontwo-dimensional gel electrophoresis profiles were analyzed to find out the differenceform the five groups protein expression.3.With Mascot2.2software retrieves the corresponding database to find out thedifferences in protein expression point, matrix-assisted laser desorption ionizationtime of flight mass spectrometry (MALDI-TOF/TOF-MS) measured peptide massfingerprint, retrieve Swiss-Port database, for differentially expressed protein spotswere identified, the results obtained protein identification.Results：1. Normal group compared with model group：The disappearance ofprotein:14-3-3protein eta, sirtuin (silent mating type information regulation2 homolog)2(s.cerevisiae isoform CRA-b); downregulated proteins: tropomyosinalpha-3,NADH dehydrogenase [ubiquinone] iron-sulfur protein2(reducednicotinamide adenine dinucleotide dehydrogenase).2.Model acupuncture group compared with model group: emergingprotein:14-3-3protein eta, sirtuin (silent mating type information regulation2homolog)2(s.cerevisiae isoform CRA-b); upregulation of proteins: tropomyosinalpha-3, microtubule-associated protein-2kinase, NADH dehydrogenase [ubiquinone]iron-sulfur protein2(reduced nicotinamide gland purine dinucleotide dehydrogenase).3.Bundled with the blank control group compared with normal control group: thedisappearance of proteins: tropomyosin alpha-3, NADH dehydrogenase [ubiquinone]iron-sulfur protein2(reduced nicotinamide adenine dinucleotide dehydrogenase);protein emerging: microtubule-associated protein-2kinase.4.Bundled with the normal control group compare with model group: thedisappearance of protein:14-3-3protein eta, sirtuin (silent mating type informationregulation2homolog)2(s.cerevisiae isoform CRA-b); expression down proteins:microtubule-associated protein-2kinase.Conclusion：1.Application of proteomics technology, discovered proteinsassociated with insomnia, as well as the efficacy of acupuncture treatment ofinsomnia-related proteins.2.The use of proteins associated with insomnia, for help further study insomniadisease markers and drug targets; utilization and efficacy of acupuncture treatment ofinsomnia related proteins, can further explore the mechanism of acupuncturetreatment for insomnia molecular biology.