Comparative Proteome Analysis In Hippocampus And Prefrontal Cortex Underling Extinction And Reinstatement Of Morphine Dependence

Objective To use comparative proteomics technology to find out the different expressed protein in HIP and PFC between extinction and reinstatement of morphine addiction.To find out special proteins of different states and find critical proteins in the reinstatement.The outcome may give us a clue to uncover the complex mechanism of morphine dependence.Drug Treatment of dependence may be uncovered to the protein.Methods 60 male SD rat were tested randomly in 3 groups:saline group morphine group and yoked morphine group.The morphine reinstatement model was established by vein self-administration.Effective nose pokes was counted as main index.After anesthesia,the hippocampus were separated by the rat stereo taxis instrument on ice.The separated tissue was put into liquid nitrogen for 5min,then store in the -80℃fridge。Extract and purify the total protein of the hippocampus tissue.The immobilized pH gradient 2-dimentional polyacrylamide gel electrophoresis(2-DE) was adopted to separate the proteins in 6 groups. After the silver staining,the PDQuest 2-DE software was applied to analyze the image.Peptid dactylograms were matched in protein data bank on internet.Result①the The morphine group active pokes significantly increased after 13days of training,compared with the saline group (P＜0.001).After 12 days withdrawal training,The yoked morphine group significantly increased compared withdrawal period(p＜0.001).T he saline group was also increased but insignificantly.②In the hippocampus 11 kinds of protein were found.They Are cAMP-dependent protein kinase inhibitor gamma,Fatty acid-biding protein,Protein p8 MTCP-1,Nucleoside diphosphate kinase A,EF-hand calcium-binding domain-contain protein 1,K channel-interacting protein 2,NADH dehydrogenase[ubiquinone]1 alpha subcomplex subunit 10,Carboxylesterase 3,Cytoskeleton-associated protein 4,Translation initiation factor eIF-2B subunit alpha and Heat shock-related 70 kDa protein 2③In the Prefrontal cortex 13 kinds of protein were found.They are Cell division cycle-associated protein 3, Protein phosphates 1 regulatory subunit 7,Heat shock 70 kDa protein 14, UMP-CMP kinase,Ubiquitin-associated protein 1,Glucokinase,26S protease regulatory subunit 7,Proteasome subunit alpha type-6 Cytoskeleton-associated protein 4,they are Endophilin-A2,NADH dehydrogenase[ubiquinone]1 alpha subcomplex subunit 10 and KIF1-binding protein. Conclusion①In this study,the morphine extinction and reinstatement modals were successfully established by self-admistration.The modals are partly same to human drug taking modals.they are real stably drug dependence modals.②In this study,proteomics technologies were successfully performed in the morphine dependence modals.In the hippocampus 4 main kinds of protein are analysised.In the Prefrontal cortex 3 main kinds of protein are analysised.They mainly plays important parts in energy metabolism,cell reconstitution and cell differentiation They may provide scientific foundation for the screening the molecular biomarker used to diagnose and treat opium dependence,as well as to provide new clue for research of drug addiction.