To screen the differential protein of ACC brain region in pain memory model rats by proteomics method，to further screen the differential proteins related to electroacupuncture intervention pain memory，so as to provide theoretical support for the further study of electroacupuncture intervention pain memory.Methods:A total of 18 healthy male Sprague Dawley rats were divided into three groups，with 6 rats in each group，including a control group，a model group and an EA group.The pain memory model was made by twice cross-injection carrageenan in EA and model groups.The control group was given the same dose of normal saline in the same time point as the model and EA group.EA was applied to bilateral Zusanli(ST36)and given at 5 h，1～5 days after the first injection for 30 min per time.The model group was taken the same measures except for EA.The pain withdrawal thresholds were detected before modeling and at 4 h，120 h after the first injection and before second injection and at 4 h，72 h after the second injection.Rats were killed at 3rd day after the second injection and taking the ACC of rats for preparation.Samples were separated by 2-D gel electrophoresis(2-DE).Then we got different samples of proteomic map by different excitation wavelength light scan.The gels were respectively image analyzed by Image Master 2D 60，to screen the protein changed in abundance more than 15 folder up or down compared with which in constitution of the control group as differential proteomics，and then had the mass spectrum identification.Results:1)Compared with the blank group，the pain threshold of carrageenan ankle was significantly decreased in the model group(P<005).Compared with the model group，the ankle pain threshold of electroacupuncture group without twice cross-injection carrageenan was rised significantly(P<005).)2)After proteomic analysis，18 differential proteins were screened out.Among them，there were 11 with significant difference in the model group，4 of them had down expression，including Tubulin alpha-1C chain，DAB2 interacting protein，NADH dehydrogenase ［ubiquinone］ flavoprotein 2，Transglutinin-3; and 7 of them had up expression，including Tubulin beta-3，Phosphoglycerate kinase 1，Steroidogenic acute regulatory protein，Actin cytoplasmic 2，Cytochrome c oxidase subunit 6A1，Ubiquitin-40S ribosomal protein S27a.Compared with the blank group，EA group had 15 proteins with significant differences，3 of them had down expression，including Tubulin alpha-1C chain，Rho GDP-dissociation inhibitor 1，NADH dehydrogenase ［ubiquinone］ flavoprotein 2，and 12 of them had up expression，including Tubulin beta-2A chain，Tubulin beta-3A chain，Actin 1，aconitate hydratase，pyruvate kinase isoenzyme，isocitrate dehydrogenase，phosphoglycerate kinase 1，Ras-related Rab-19 protein，steroid hormone synthesis of acute protein，actin 2，cells Pigment c oxidase 6A1 subunit，ubiquitin 40S ribosomal protein S27a.There were 8 differentially expressed proteins in the EA group compared with the model group，and 2 were down-regulated，which were Rho GDP dissociation inhibitor and actin 26 were up-regulated，respectively，tubulin α-1A Chain，tubulin β-2A chain，DAB2 interacting protein，aconitate hydratase，isocitrate dehydrogenase，Ras-related Rab-19 protein.Conclusion:There were 11 differential proteomics involved in pain memory of awakening process through proteomics analysis.And 8 differential proteomics were involved in the intervention of EA on pain memory.The intervention effect of electroacupuncture(EA)pretreatment on pain memory may be related to inhibition of synaptic plasticity.And the inhibition may be related to reinforce nerve cytoskeleton structure.