Cellular and Molecular Neurobiology

, Volume 37, Issue 4, pp 643–653 | Cite as

Involvement of Spinal PKMζ Expression and Phosphorylation in Remifentanil-Induced Long-Term Hyperalgesia in Rats

  • Qi Zhao
  • Linlin Zhang
  • Ruichen Shu
  • Chunyan Wang
  • Yonghao Yu
  • Haiyun Wang
  • Guolin Wang
Original Research


Up-regulation of GluN2B-containing N-methyl-d-aspartate receptors (NMDARs) expression and trafficking is the key mechanism for remifentanil-induced hyperalgesia (RIH), nevertheless, the signaling pathway and pivotal proteins involved in RIH remain equivocal. PKMζ, an isoform of protein kinase C (PKC), maintains pain memory storage in neuropathic pain and inflammatory pain, which plays a parallel role regulated by NMDARs in long-term memory trace. In the present study, Zeta Inhibitory Peptide (ZIP), a PKMζ inhibitor, and a selective GluN2B antagonist Ro-256981 are injected intrathecally before remifentanil infusion (1 μg kg−1 min−1 for 1 h, iv) in order to detect whether GluN2B contributes to RIH through affecting synthesis and activity of PKMζ in spinal dorsal horn. Nociceptive tests are measured by Paw withdrawal mechanical threshold (PWT) and paw withdrawal thermal latency (PWL). The L4–L6 segments of dorsal horn taken from rats with RIH are for determining expression of PKMζ and pPKMζ by Western blot and immunohistochemistry. Our data suggest that remifentanil infusion causes an increase of PKMζ in expression and phosphorylation in rats with nociceptive sensitization, beginning at 2 h, peaked at 2 days, and returned to basal level at 7 days. ZIP (10 ng) could block behavioral sensitization induced by remifentanil. Ro25-6981 dosage-dependently attenuated mechanical and thermal hyperalgesia and reversed expression of PKMζ and pPKMζ, indicating that GluN2B-containing NMDA receptor facilitates development of RIH through mediating expression and activity of spinal PKMζ in rats. Although detailed mechanisms require further comprehensive study, the preventive role of Ro25-6981 and ZIP provide novel options for the effective precaution of RIH in clinics.


Hyperalgesia PKMζ ZIP N-methyl-d-aspartate Remifentanil 



N-methyl-d-aspartate receptors


Remifentanil-induced hyperalgesia


Protein kinase C


Zeta inhibitory peptide


Paw withdrawal mechanical threshold


Paw withdrawal thermal latency



This work was supported by research grants from the National Natural Science Foundation of China (81571077, 81371245, 81301025, and 81400908) and Science and Technology Supported Key Project of Tianjin (12ZCZDSY03000).

Author contribution

Qi Zhao helped design the study, conduct the study, analyze the data, and write the manuscript. Linlin Zhang helped design the study, conduct the study, and analyze the data. Ruichen Shu helped conduct the study and analyze the data. Chunyan Wang and Haiyun Wang helped design the study and analyze the data. Yonghao Yu helped conduct the study. Guolin Wang helped design the study, analyze the data, and write the manuscript.

Compliance with Ethical Standards

Conflict of Interest

The authors declare no conflict of interest.


  1. Andre L, Mark HP, Anne H, Yue H, Virginia C (2011) PKM ζ is essential for spinal plasticity underlying the maintenance of persistent pain. Mol Pain 7:99Google Scholar
  2. Angst MS, Koppert W, Pahl I, Clark DJ, Schmelz M (2003) Short-term infusion of mu-opioid agonist remifentanil in humans causes hyperalgesia during withdrawal. Pain 106:49–57CrossRefPubMedGoogle Scholar
  3. Asiedu MN, Tillu DV, Melemedjian OK, Shy A, Sanoja R, Bodell B, Ghosh S, Porreca F, Price TJ (2011) Spinal protein kinase M zeta underlies the maintenance mechanism of persistent nociceptive sensitization. J Neurosci 31(18):6646–6653CrossRefPubMedPubMedCentralGoogle Scholar
  4. Bornemann-Cimenti H, Lederer AJ, Wejbora M, Michaeli K, Kern-Pirsch C, Archan S (2012) Preoperative pregabalin administration significantly reduces postoperative opioid consumption and mechanical hyperalgesia after transperitoneal nephrectomy. Br J Anaesth 108:845–849CrossRefPubMedGoogle Scholar
  5. Celerier E, Gonzalez JR, Maldonado R, Cabañero D (2006) Opioid-induced hyperalgesia in a murine model of postoperative pain. Anesthesiology 104:546–555CrossRefPubMedGoogle Scholar
  6. Céline H, Ruth DS, Dimitris NX, Jürgen S (2011) Distinct mechanisms underlying pronociceptive effects of opioids. J Neurosci 31(46):16748–16756CrossRefGoogle Scholar
  7. Chizh BA (2007) Low dose ketamine: a therapeutic and research tool to explore N-methyl-d-aspartate (NMDA) receptor-mediated plasticity in pain pathways. J Psychopharmacol 21:259–271CrossRefPubMedGoogle Scholar
  8. Chu LF, Angst MS, Clark D (2008) Opioid-induced hyperalgesia in humans: molecular mechanisms and clinical considerations. Clin J Pain 24:479–496CrossRefPubMedGoogle Scholar
  9. Crain SM, Shen KF (2000) Antagonists of excitatory opioid receptor functions enhance morphine’s analgesic potency and attenuate opioid tolerance/dependence liability. Pain 84:121–131CrossRefPubMedGoogle Scholar
  10. Derrode N, Lebrun F, Levron JC, Chauvin M, Debaene B (2003a) Influence of reoperative opioid on postoperative pain after major abdominal surgery: sufentanil TCI versus remifentanil TCI. A randomized, controlled study. Br J Anaesth 91:842–849CrossRefPubMedGoogle Scholar
  11. Derrode N, Lebrun F, Levron JC, Chauvin M, Debaene B (2003b) Influence of preoperative opioid on postoperative pain after major abdominal surgery: sufentanil TCI versus remifentanil TCI. A randomized, controlled study. Br J Anaesth 93:409–417Google Scholar
  12. Drdla R, Gassner M, Gingl E,Open image in new window J (2009) Induction of synaptic long-term potentiation after opioid withdrawal. Science 325(5937):207–210CrossRefPubMedGoogle Scholar
  13. Gu X, Wu X, Liu Y, Cui S, Ma Z (2009) Tyrosine phosphorylation of the N-methyl-d-aspartate receptor 2B subunit in spinal cord contributes to remifentanil-induced postoperative hyperalgesia: the preventive effect of ketamine. Mol Pain 5:76CrossRefPubMedPubMedCentralGoogle Scholar
  14. Jiang M, Zhang W, Ma ZL, Gu XP (2013) Antinociception and prevention of hyperalgesia by intrathecal administration of Ro 25-6981, a highly selective antagonist of the 2B subunit of N-methyl-d-aspartate receptor. Pharmacol Biochem Behav 112:56–63CrossRefPubMedGoogle Scholar
  15. Kelly MT, Crary JF, Scaktor TC (2007) Regulation of protein kinase Mzeta synthesis by multiple kinases in long-term potentiation. J Neurosci 27:3439–3444CrossRefPubMedGoogle Scholar
  16. King T, Qu C, Okun A, Melemedjian OK, Mandell EK, Maskaykina IY, Navratilova E, Dussor GO, Ghosh S, Price TJ, Porreca F (2012) Contribution of PKM-zeta dependent and independent amplification to components of experimental neuropathic pain. Pain 153(6):1263–1273CrossRefPubMedPubMedCentralGoogle Scholar
  17. Kissin I, Lee SS, Arthur GR, Bradley EJ (1996) Time course characteristics of acute tolerance development to continuously infused alfentanial in rats. Anesth Analg 83:600–605CrossRefPubMedGoogle Scholar
  18. Klaus H, Joke N, Hugo K, Buerkle H, Halene T, Schauerte S (2004) Remifentanil directly activates human N-methyl-d-aspartate receptors expressed in Xenopus laevis oocytes. Anesthesiology 100:1531–1537CrossRefGoogle Scholar
  19. Koppert W, Schmelz M (2007) The impact of opioid-induced hyperalgesia for postoperative pain. Best Pract Res Clin Anaesthesiol 21:65–83CrossRefPubMedGoogle Scholar
  20. Lee M, Silverman SM, Hansen H, Patel VB, Manchikanti L (2011) A comprehensive review of opioid-induced hyperalgesia. Pain Physician 14:145–161PubMedGoogle Scholar
  21. Li XY, Ko HG, Chen T (2010) Alleviating neuropathic pain hypersensitivity by inhibiting PKM zeta in the anterior ingulated cortex. Science 330(6009):1400–1404CrossRefPubMedGoogle Scholar
  22. Li Y, Wang H, Xie K, Wang C, Yang Z, Yu Y, Wang G (2013) Inhibition of glycogen synthase kinase-3β prevents remifentanil-induced hyperalgesia via regulating the expression and function of spinal N-methyl-d-aspartate receptors in vivo and vitro. PLoS ONE 8(10):e77790CrossRefPubMedPubMedCentralGoogle Scholar
  23. Liang DY, Li X, Clark JD (2011) 5-Hydroxytryptamine type 3 receptor modulates opioid-induced hyperalgesia and tolerance in mice. Anesthesiology 114:1180–1189CrossRefPubMedPubMedCentralGoogle Scholar
  24. Rivosecchi RM, Rice MJ, Smithburger PL, Buckley MS, Coons JC, Kane-Gill SL (2014) An evidence based systematic review of remifentanil associated opioid-induced hyperalgesia. Expert Opin Drug Saf 13(5):587–603CrossRefPubMedGoogle Scholar
  25. Sacktor TC (2011) How does PKM ζ maintain long-term memory? Nat Rev Neurosci 12(1):9–15CrossRefPubMedGoogle Scholar
  26. Sadeh N, Verbitsky S, Dudai Y, Seqal M (2015) Zeta inhibitory peptide, a candidate inhibitor of protein kinase Mζ, is excitotoxic to cultured hippocampal neurons. J Neurosci 35(36):12404–12411CrossRefPubMedGoogle Scholar
  27. Shema R, Haramati S, Ron S, Hazvi S, Chen A, Sacktor TC, Dudai Y (2011) Enhancement of consolidated long-term memory by overexpression of protein kinase Mζ in the neocortex. Science 331(6021):1207–1210CrossRefPubMedGoogle Scholar
  28. Størkson RV, Kjørsvik A, Tjølsen A, Hole K (1996) Lumbar catheterization of the spinal subarachnoid space in the rat. J Neurosci Methods 65:167–172CrossRefPubMedGoogle Scholar
  29. Sullivan JP, Connor JR, Shearer BG, Burch RM (2002) 2,6-Diamino-N-([1-(1oxotridecyl)-2-piperidinyl] methyl)hexanamide (NPC 15437), a novel inhibitor of protein kinase C interacting at the regulatory domain. Mol Pharmacol 41:38–44Google Scholar
  30. Todd CS (2012) Memory maintenance by PKM ζ-an evolutionary perspective. Sacktor Mol Brain 5:31CrossRefGoogle Scholar
  31. Von Engelhardt J, Coserea I, Pawlak V, Fuchs EC, Seeburg PH, Monyer H (2007) Excitotoxicity in vitro by NR2A- and GLUN2B-containing NMDA receptors. Neuropharmacology 53:10–17CrossRefGoogle Scholar
  32. Wider-smith OH, Arendt-Nielsen L (2006) Postoperative hyperalgesia: its clinical importance and relevance. Anesthesiology 104:106–107Google Scholar
  33. Yuan Y, Wang JY, Yuan F, Xie KL, Yu YH, Wang GL (2013) Glycogen synthase kinase-3β contributes to remifentanil-induced postoperative hyperalgesia via regulating N-methyl-d-aspartate receptor trafficking. Anesth Analg 116(2):473–481CrossRefPubMedGoogle Scholar
  34. Zhang LL, Shu RC, Wang HY, Yu YH, Wang CY, Yang M, Wang M, Wang GL (2014) Hydrogen-rich saline prevents remifentanil-induced hyperalgesia and inhibits MnSOD nitration via regulation of GLUN2B-containing NMDA receptors in rats. Neuroscience 280:171–180CrossRefPubMedGoogle Scholar
  35. Zhao M, Joo DT (2008a) Enhancement of spinal N-methyl-d-aspartate receptor function by remifentanil action at delta-opioid receptors as a mechanism for acute opioid-induced hyperalgesia or tolerance. Anesthesiology 109(2):308–317CrossRefPubMedGoogle Scholar
  36. Zhao M, Joo DT (2008b) Enhancement of spinal N-methyl-D-aspartate receptor function by remifentanil action at delta-opioid receptors as a mechanism for acute opioid-induced hyperalgesia or tolerance. Anesthesiology 109(2):308–317CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Qi Zhao
    • 1
    • 2
  • Linlin Zhang
    • 1
    • 2
  • Ruichen Shu
    • 1
    • 2
  • Chunyan Wang
    • 1
    • 2
  • Yonghao Yu
    • 1
    • 2
  • Haiyun Wang
    • 1
    • 2
  • Guolin Wang
    • 1
    • 2
  1. 1.Department of AnesthesiologyTianjin Medical University General HospitalTianjinChina
  2. 2.Tianjin Research Institute of AnesthesiologyTianjinChina

Personalised recommendations