title: Expression and regulation of neurotrophic and angiogenic factors during human intervertebral disc degeneration.
abstract: BACKGROUND
  The degenerate intervertebral disc (IVD) becomes innervated by sensory nerve fibres, and vascularised by blood vessels. This study aimed to identify neurotrophins, neuropeptides and angiogenic factors within native IVD tissue and to further investigate whether pro-inflammatory cytokines are involved in the regulation of expression levels within nucleus pulposus (NP) cells, nerve and endothelial cells.
  
  
  METHODS
  Quantitative real-time PCR (qRT-PCR) was performed on 53 human IVDs from 52 individuals to investigate native gene expression of neurotrophic factors and their receptors, neuropeptides and angiogenic factors. The regulation of these factors by cytokines was investigated in NP cells in alginate culture, and nerve and endothelial cells in monolayer using RT-PCR and substance P (SP) protein expression in interleukin-1 (IL-1β) stimulated NP cells.
  
  
  RESULTS
  Initial investigation on uncultured NP cells identified expression of all neurotrophins by native NP cells, whilst the nerve growth factor (NGF) receptor was only identified in severely degenerate and infiltrated discs, and brain derived neurotrophic factor (BDNF) receptor expressed by more degenerate discs. BDNF expression was significantly increased in infiltrated and degenerate samples. SP and vascular endothelial growth factor (VEGF) were higher in infiltrated samples. In vitro stimulation by IL-1β induced NGF in NP cells. Neurotropin-3 was induced by tumour necrosis factor alpha in human dermal microvascular endothelial cells (HDMECs). SP gene and protein expression was increased in NP cells by IL-1β. Calcitonin gene related peptide was increased in SH-SY5Y cells upon cytokine stimulation. VEGF was induced by IL-1β and interleukin-6 in NP cells, whilst pleiotrophin was decreased by IL-1β. VEGF and pleiotrophin were expressed by SH-SY5Y cells, and VEGF by HDMECs, but were not modulated by cytokines.
  
  
  CONCLUSIONS
  The release of cytokines, in particular IL-1β during IVD degeneration, induced significant increases in NGF and VEGF which could promote neuronal and vascular ingrowth. SP which is released into the matrix could potentially up regulate the production of matrix degrading enzymes and also sensitise nerves, resulting in nociceptive transmission and chronic low back pain. This suggests that IL-1β is a key regulatory cytokine, involved in the up regulation of factors involved in innervation and vascularisation of tissues.
authors.forename: Abbie L A
authors.initials: AL
authors.lastname: Binch
authors.forename: Ashley A
authors.initials: AA
authors.lastname: Cole
authors.forename: Lee M
authors.initials: LM
authors.lastname: Breakwell
authors.forename: Anthony L R
authors.initials: AL
authors.lastname: Michael
authors.forename: Neil
authors.initials: N
authors.lastname: Chiverton
authors.forename: Alison K
authors.initials: AK
authors.lastname: Cross
authors.forename: Christine L
authors.initials: CL
authors.lastname: Le Maitre
mesh_terms: D000328:Adult; D000368:Aged; D043925:Angiogenesis Inducing Agents / Q000378:metabolism*; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics; D015740:Calcitonin Gene-Related Peptide / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007403:Intervertebral Disc / Q000378:metabolism* / Q000473:pathology; D055959:Intervertebral Disc Degeneration / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D020932:Nerve Growth Factor / Q000235:genetics; D009414:Nerve Growth Factors / Q000235:genetics*; D017475:Receptors, Nerve Growth Factor / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013373:Substance P / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics; D055815:Young Adult
chemical_list: D043925:Angiogenesis Inducing Agents; D019208:Brain-Derived Neurotrophic Factor; D002352:Carrier Proteins; D016207:Cytokines; D009414:Nerve Growth Factors; D017475:Receptors, Nerve Growth Factor; D042461:Vascular Endothelial Growth Factor A; C066845:pleiotrophin; D013373:Substance P; D020932:Nerve Growth Factor; D015740:Calcitonin Gene-Related Peptide