title: Disk injury in rats produces persistent increases in pain-related neuropeptides in dorsal root ganglia and spinal cord glia but only transient increases in inflammatory mediators: pathomechanism of chronic diskogenic low back pain.
abstract: METHODS
  Immunohistological analysis in an injured intervertebral disk (IVD) model.
  
  
  OBJECTIVE
  To elucidate and compare in rats the behavior of the sensory nervous system and inflammatory mediators in experimentally injured IVDs.
  
  
  BACKGROUND
  Multiple human and animal studies have verified the presence of sensory nerve fibers in IVDs or investigated the behavior of inflammatory mediators in injured IVDs, but no in vivo study to date has examined the relationship between the 2.
  
  
  METHODS
  Eight-week-old female rats were used. In the disk-injured group, L5/L6 disks were injured with a 24-gauge needle; simultaneously, the neurotracer Fluoro-gold was injected into the L5/L6 IVD. The L5/L6 IVD dorsal root ganglia (DRGs) from the L1 to L6 levels, and the spinal cord was resected at several time points after surgery. Nerve growth factor, tumor necrosis factor (TNF)-α and interleukin (IL)-6 production in the IVDs were quantified using enzyme-linked immunosorbent assay. DRGs were immunostained for calcitonin gene-related peptide, and spinal cord sections were immunostained for ionized calcium-binding adaptor molecule-1 and glial fibrillary acidic protein.
  
  
  RESULTS
  Nerve growth factor, and TNF-α levels (through 1 week) and IL-6 levels (through 4 days) were significantly higher in the disk-injured group than in the noninjured group (P < 0.05). However, starting at 2 weeks (nerve growth factor and TNF-α) or 1 week (IL-6), the differences in inflammatory mediator levels between the 2 groups no longer were significant. In contrast, the percentage of calcitonin gene-related peptide-immunoreactive neurons among Fluoro-gold-labeled DRG neurons, and the numbers of ionized calcium-binding adaptor molecule-1-immunoreactive microglia and glial fibrillary acidic protein-immunoreactive astrocytes in the spinal dorsal horn remained significantly higher in the injured group than in the noninjured group at all-time points (P < 0.05).
  
  
  CONCLUSIONS
  Disk injury in rats produces persistent increases in neuropeptides in DRGs and glia in the spinal cord, but only transient increases in inflammatory mediators in IVDs.
authors.affiliation: Department of Orthopaedic Surgery, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba, Japan. masayuki008@aol.com
authors.forename: Masayuki
authors.initials: M
authors.lastname: Miyagi
authors.forename: Tetsuhiro
authors.initials: T
authors.lastname: Ishikawa
authors.forename: Sumihisa
authors.initials: S
authors.lastname: Orita
authors.forename: Yawara
authors.initials: Y
authors.lastname: Eguchi
authors.forename: Hiroto
authors.initials: H
authors.lastname: Kamoda
authors.forename: Gen
authors.initials: G
authors.lastname: Arai
authors.forename: Miyako
authors.initials: M
authors.lastname: Suzuki
authors.forename: Gen
authors.initials: G
authors.lastname: Inoue
authors.forename: Yasuchika
authors.initials: Y
authors.lastname: Aoki
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Toyone
authors.forename: Kazuhisa
authors.initials: K
authors.lastname: Takahashi
authors.forename: Seiji
authors.initials: S
authors.lastname: Ohtori
mesh_terms: D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005727:Ganglia, Spinal / Q000378:metabolism*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D018836:Inflammation Mediators / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism; D007403:Intervertebral Disc / Q000293:injuries*; D008159:Lumbar Vertebrae / Q000293:injuries*; D008840:Microfilament Proteins / Q000378:metabolism; D020932:Nerve Growth Factor / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism*; D009479:Neuropeptides / Q000378:metabolism*; D010146:Pain / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: C100745:Aif1 protein, rat; D002135:Calcium-Binding Proteins; D005904:Glial Fibrillary Acidic Protein; D018836:Inflammation Mediators; D015850:Interleukin-6; D008840:Microfilament Proteins; D009479:Neuropeptides; D014409:Tumor Necrosis Factor-alpha; D020932:Nerve Growth Factor; D015740:Calcitonin Gene-Related Peptide