title: CC chemokine ligand 2 upregulates the current density and expression of TRPV1 channels and Nav1.8 sodium channels in dorsal root ganglion neurons.
abstract: BACKGROUND
  Inflammation or nerve injury-induced upregulation and release of chemokine CC chemokine ligand 2 (CCL2) within the dorsal root ganglion (DRG) is believed to enhance the activity of DRG nociceptive neurons and cause hyperalgesia. Transient receptor potential vanilloid receptor 1 (TRPV1) and tetrodotoxin (TTX)-resistant Na(v)1.8 sodium channels play an essential role in regulating the excitability and pain transmission of DRG nociceptive neurons. We therefore tested the hypothesis that CCL2 causes peripheral sensitization of nociceptive DRG neurons by upregulating the function and expression of TRPV1 and Nav1.8 channels.
  
  
  METHODS
  DRG neuronal culture was prepared from 3-week-old Sprague-Dawley rats and incubated with various concentrations of CCL2 for 24 to 36 hours. Whole-cell voltage-clamp recordings were performed to record TRPV1 agonist capsaicin-evoked inward currents or TTX-insensitive Na(+) currents from control or CCL2-treated small DRG sensory neurons. The CCL2 effect on the mRNA expression of TRPV1 or Na(v)1.8 was measured by real-time quantitative RT-PCR assay.
  
  
  RESULTS
  Pretreatment of CCL2 for 24 to 36 hours dose-dependently (EC(50) value = 0.6 ± 0.05 nM) increased the density of capsaicin-induced currents in small putative DRG nociceptive neurons. TRPV1 mRNA expression was greatly upregulated in DRG neurons preincubated with 5 nM CCL2. Pretreating small DRG sensory neurons with CCL2 also increased the density of TTX-resistant Na(+) currents with a concentration-dependent manner (EC(50) value = 0.7 ± 0.06 nM). The Na(v)1.8 mRNA level was significantly increased in DRG neurons pretreated with CCL2. In contrast, CCL2 preincubation failed to affect the mRNA level of TTX-resistant Nav1.9. In the presence of the specific phosphatidylinositol-3 kinase (PI3K) inhibitor LY294002 or Akt inhibitor IV, CCL2 pretreatment failed to increase the current density of capsaicin-evoked inward currents or TTX-insensitive Na(+) currents and the mRNA level of TRPV1 or Na(v)1.8.
  
  
  CONCLUSIONS
  Our results showed that CCL2 increased the function and mRNA level of TRPV1 channels and Na(v)1.8 sodium channels in small DRG sensory neurons via activating the PI3K/Akt signaling pathway. These findings suggest that following tissue inflammation or peripheral nerve injury, upregulation and release of CCL2 within the DRG could facilitate pain transmission mediated by nociceptive DRG neurons and could induce hyperalgesia by upregulating the expression and function of TRPV1 and Na(v)1.8 channels in DRG nociceptive neurons.
authors.affiliation: Department of Physiology and Pharmacology, Chang Gung University School of Medicine, Kwei-San, Tao-Yuan 333, Taiwan.
authors.forename: Der-Jang
authors.initials: DJ
authors.lastname: Kao
authors.forename: Allen H
authors.initials: AH
authors.lastname: Li
authors.forename: Jin-Chung
authors.initials: JC
authors.lastname: Chen
authors.forename: Ro-Sun
authors.initials: RS
authors.lastname: Luo
authors.forename: Ying-Ling
authors.initials: YL
authors.lastname: Chen
authors.forename: Juu-Chin
authors.initials: JC
authors.lastname: Lu
authors.forename: Hung-Li
authors.initials: HL
authors.lastname: Wang
mesh_terms: D000200:Action Potentials / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000502:physiology*; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism*; D062559:NAV1.8 Voltage-Gated Sodium Channel / Q000096:biosynthesis* / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D050916:TRPV Cation Channels / Q000096:biosynthesis* / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics*
chemical_list: C491766:Ccl2 protein, rat; D018932:Chemokine CCL2; D062559:NAV1.8 Voltage-Gated Sodium Channel; D012333:RNA, Messenger; C568319:Scn10a protein, rat; D050916:TRPV Cation Channels; C493922:Trpv1 protein, rat