title: Identification of differentially expressed proteins in the spinal cord of neuropathic pain models with PKCgamma silence by proteomic analysis.
abstract: In order to elucidate the mechanisms that PKCγ regulates neuropathic pain (NP), and detect proteins that are associated with the function of PKCγ in NP, we exploited a chronic constriction injury (CCI)-induced neuropathic pain rat (CCI-NP rat) model in which PKCγ knockdown in the spinal cord was successfully carried out with stable RNA interference (RNAi). The spinal cords (L4-L5) were surgically obtained from CCI-NP rats with and without PKCγ knockdown, for comparative proteomic analysis. The total proteins from the spinal cords (L4-L5) were extracted and were separated with two-dimensional gel electrophoresis (2DGE). 2D gel images were analyzed with PDQuest software. Nineteen differential gel-spots were identified with spot-volume increased and 17 spots with spot-volume decreased. Among them, eighteen differentially expressed proteins (DEPs) were identified with matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) between CCI-NP rats with and without PKCγ knockout. Those DEPs are involved in transmission and modulation of noxious information; cellular homeostasis and metabolism; antioxidant proteins, heat shock proteins and chaperones; membrane receptor trafficking; and cytoskeleton. Three DEPs (SNAP-25, TERA and AR) were validated with Western blot analysis, and confirmed the DEP data. Further study showed that AR-selective inhibitor epalrestat totally turned over the upregulated expression of AR in CCI-NP rats. Those DEP data are extensively associated with the function of PKCγ that regulates NP, and would contribute to the clarification of the mechanisms of PKCγ in NP.
authors.affiliation: Department of Anesthesiology, Xiangya Hospital, Central South University, Changsha, Hunan 410008, China.
authors.forename: Wangyuan
authors.initials: W
authors.lastname: Zou
authors.forename: Xianquan
authors.initials: X
authors.lastname: Zhan
authors.forename: Maoyu
authors.initials: M
authors.lastname: Li
authors.forename: Zongbin
authors.initials: Z
authors.lastname: Song
authors.forename: Chang
authors.initials: C
authors.lastname: Liu
authors.forename: Fang
authors.initials: F
authors.lastname: Peng
authors.forename: Qulian
authors.initials: Q
authors.lastname: Guo
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000449:Aldehyde Reductase / Q000378:metabolism; D000818:Animals; D003251:Constriction, Pathologic; D015180:Electrophoresis, Gel, Two-Dimensional; D007150:Immunohistochemistry; D008297:Male; D009437:Neuralgia / Q000378:metabolism*; D011493:Protein Kinase C / Q000235:genetics* / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D040901:Proteomics; D034622:RNA Interference; D051381:Rats; D017207:Rats, Sprague-Dawley; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013116:Spinal Cord / Q000378:metabolism*; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism
chemical_list: D011506:Proteins; C494210:Snap25 protein, rat; D050825:Synaptosomal-Associated Protein 25; D000449:Aldehyde Reductase; C087857:protein kinase C gamma; D011493:Protein Kinase C; D000251:Adenosine Triphosphatases