Expression of TRPV1 and iNOS in the Dorsal Root Ganglion Exposed by Autologous Nucleus Pulposus in the Rat

  • Kim, Su-Jeong (Institute of Medical Science, Yeungnam University) ;
  • Seo, Jeong-Min (Institute of Biomedical Engineering, Yeungnam University) ;
  • Cho, Yun-Woo (Department of Rehabilitation Medicine, College of Medicine, Yeungnam University) ;
  • Park, Hea-Woon (Department of Rehabilitation Medicine, School of Medicine, Catholic University of Deagu) ;
  • Lee, Joon-Ha (Department of Biochemistry and Molecular Biology, Yeungnam University) ;
  • Hwang, Se-Jin (Department of Anatomy and Cell Biology, College of Medicine, Hanyang University) ;
  • Ahn, Sang-Ho (Department of Rehabilitation Medicine, College of Medicine, Yeungnam University)
  • Received : 2010.05.13
  • Accepted : 2010.06.14
  • Published : 2010.06.25

Abstract

Purpose: To determine whether upregulation of inducible nitric oxide synthase (iNOS) transcription and translation is related to radicular pain in a model of lumbar disc herniation. Also, to investigate the temporal changes of mRNA expression of iNOS and the identity of iNOS and transient receptor potential vanilloid (TRPV) 1 channel expression cells in dorsal root ganglion (DRG) of a model of lumbar disc herniation. Methods: A lumbar disc herniated rat model was developed by implantation of the autologous nucleus pulposus, harvested from the coccygeal vertebra of each tail, on the left L5 nerve root just proximal to the DRG. Rats were tested for mechanical allodynia of the plantar surface of both hind paws 2 days before surgery and 1, 5, 10, 20 and 30 days postoperatively. Reverse transcription polymerase chain reaction (RT-PCR) was used to follow iNOS mRNA expression. To stain iNOS and TRPV1 in DRG, an immunohistochemical study was done 10 days after surgery. Results: A significant drop in mechanical withdrawal threshold on the ipsilateral and contralateral hind paws was observed 1 day after surgery and was prolonged to 30 days in rats with lumbar disc herniation. The expression of mRNA for iNOS peaked at postoperative day 10 on both sides of the DRG. iNOS-positive sensory neurons in the DRG varied in size from large to small diameter cells. A majority of small and intermediate sensory neurons were TRPV1-positive cells. Double immunofluorescence staining for TRPV1 and iNOS revealed that most intermediate TRPV1-positive sensory neurons co-localized with iNOS-positive neurons. Conclusion: Nucleus pulposus-induced mechanical allodynia can be generated without mechanical compression. This pain is related to temporal changes in expression of iNOS mRNA in the DRG. Co-localization of TRPV1 and iNOS in intermediate neurons of the DRG is correlated with pain modality and intensity.

Keywords

References

  1. Goupille P, Mulleman D, Valat JP. Radiculopathy associated with disc herniation. Ann Rheum Dis. 2006;65(2):141-3. https://doi.org/10.1136/ard.2005.039669
  2. Olmarker K, Rydevik B, Nordborg C. Autologous nucleus pulposus induces neurophysiologic and histologic changes in porcine cauda equina nerve roots. Spine (Phila Pa 1976). 1993; 18(11):1425-32. https://doi.org/10.1097/00007632-199318110-00005
  3. Ahn SH, Cho YW, Ahn MW et al. mRNA expression of cytokines and chemokines in herniated lumbar intervertebral discs. Spine (Phila Pa 1976). 2002;27(9):911-7. https://doi.org/10.1097/00007632-200205010-00005
  4. Kawakami M, Matsumoto T, Kuribayashi K et al. mRNA expression of interleukins, phospholipase A2, and nitric oxide synthase in the nerve root and dorsal root ganglion induced by autologous nucleus pulposus in the rat. J Orthop Res. 1999;17(6):941-6. https://doi.org/10.1002/jor.1100170620
  5. Takahashi H, Suguro T, Okazima Y et al. Inflammatory cytokines in the herniated disc of the lumbar spine. Spine (Phila Pa 1976). 1996;21(2):218-24. https://doi.org/10.1097/00007632-199601150-00011
  6. Takada T, Nishida K, Doita M et al. Interleukin-6 production is upregulated by interaction between disc tissue and macrophages. Spine (Phila Pa 1976). 2004;29(10):1089-92. https://doi.org/10.1097/00007632-200405150-00007
  7. Burke JG, Watson RW, McCormack D et al. Spontaneous production of monocyte chemoattractant protein-1 and interleukin-8 by the human lumbar intervertebral disc. Spine (Phila Pa 1976). 2002;27(13):1402-7. https://doi.org/10.1097/00007632-200207010-00006
  8. Cuellar JM, Montesano PX, Carstens E. Role of TNF-alpha in sensitization of nociceptive dorsal horn neurons induced by application of nucleus pulposus to L5 dorsal root ganglion in rats. Pain. 2004;110(3):578-87. https://doi.org/10.1016/j.pain.2004.03.029
  9. Ohtori S, Takahashi K, Aoki Y et al. Spinal neural cyclooxygenase- 2 mediates pain caused in a rat model of lumbar disk herniation. J Pain. 2004;5(7):385-91. https://doi.org/10.1016/j.jpain.2004.06.004
  10. Brisby H, Byrod G, Olmarker K et al. Nitric oxide as a mediator of nucleus pulposus-induced effects on spinal nerve roots. J Orthop Res. 2000;18(5):815-20. https://doi.org/10.1002/jor.1100180520
  11. Kawakami M, Tamaki T, Hashizume H et al. The role of phospholipase A2 and nitric oxide in pain-related behavior produced by an allograft of intervertebral disc material to the sciatic nerve of the rat. Spine (Phila Pa 1976). 1997;22(10): 1074-9. https://doi.org/10.1097/00007632-199705150-00004
  12. Min SS. Effects of NOS inhibitors on arthritis and arthritic pain in rats. Korean J Physiol Pharmacol. 2007;11(6):253-7.
  13. Mandadi S, Roufogalis BD. ThermoTRP channels in nociceptors: taking a lead from capsaicin receptor TRPV1. Curr Neuropharmacol. 2008;6(1):21-38. https://doi.org/10.2174/157015908783769680
  14. Mandadi S, Numazaki M, Tominaga M et al. Activation of protein kinase C reverses capsaicin-induced calcium-dependent desensitization of TRPV1 ion channels. Cell Calcium. 2004; 35(5):471-8. https://doi.org/10.1016/j.ceca.2003.11.003
  15. Bolcskei K, Helyes Z, Szabo A et al. Investigation of the role of TRPV1 receptors in acute and chronic nociceptive processes using gene-deficient mice. Pain. 2005;117(3):368-76. https://doi.org/10.1016/j.pain.2005.06.024
  16. Christoph T, Grunweller A, Mika J et al. Silencing of vanilloid receptor TRPV1 by RNAi reduces neuropathic and visceral pain in vivo. Biochem Biophys Res Commun. 2006;350(1): 238-43. https://doi.org/10.1016/j.bbrc.2006.09.037
  17. Culshaw AJ, Bevan S, Christiansen M et al. Identification and biological characterization of 6-aryl-7-isopropylquinazolinones as novel TRPV1 antagonists that are effective in models of chronic pain. J Med Chem. 2006;49(2):471-4. https://doi.org/10.1021/jm051058x
  18. Brown BS, Keddy R, Perner RJ et al. Discovery of TRPV1 antagonist ABT-116. Bioorg Med Chem Lett. 2010;20(11): 3291-4. https://doi.org/10.1016/j.bmcl.2010.04.047
  19. Cho YW, Park HW, Kim SJ et al. Expression of MCP‐ in spinal dorsal horn in a rat model of lumbar disc herniation. J Kor Soc Phys Ther. 2009;21(3):95-102.
  20. Chaplan SR, Bach FW, Pogrel JW et al. Quantitative assessment of tactile allodynia in the rat paw. J Neurosci Methods. 1994;53(1):55-63. https://doi.org/10.1016/0165-0270(94)90144-9
  21. Sasaki N, Kikuchi S, Konno S et al. Anti-TNF-alpha antibody reduces pain-behavioral changes induced by epidural application of nucleus pulposus in a rat model depending on the timing of administration. Spine (Phila Pa 1976). 2007; 32(4): 413-6. https://doi.org/10.1097/01.brs.0000255097.18246.bc
  22. Kim SH, Chung JM. An experimental model for peripheral neuropathy produced by segmental spinal nerve ligation in the rat. Pain. 1992;50(3):355-63. https://doi.org/10.1016/0304-3959(92)90041-9
  23. Zhang JM, Li H, Brull SJ. Perfusion of the mechanically compressed lumbar ganglion with lidocaine reduces mechanical hyperalgesia and allodynia in the rat. J Neurophysiol. 2000;84(2):798-805. https://doi.org/10.1152/jn.2000.84.2.798
  24. Koltzenburg M, Wall PD, McMahon SB. Does the right side know what the left is doing? Trends Neurosci. 1999;22(3): 122-7. https://doi.org/10.1016/S0166-2236(98)01302-2
  25. Bonnefous C, Payne JE, Roppe J et al. Discovery of inducible nitric oxide synthase (iNOS) inhibitor development candidate KD7332, part 1: Identification of a novel, potent, and selective series of quinolinone iNOS dimerization inhibitors that are orally active in rodent pain models. J Med Chem. 2009;52(9):3047-62. https://doi.org/10.1021/jm900173b
  26. Kwak EK, Kim JW, Kang KS et al. The role of inducible nitric oxide synthase following spinal cord injury in rat. J Korean Med Sci. 2005;20(4):663-9. https://doi.org/10.3346/jkms.2005.20.4.663
  27. Steel JH, Terenghi G, Chung JM et al. Increased nitric oxide synthase immunoreactivity in rat dorsal root ganglia in a neuropathic pain model. Neurosci Lett. 1994;169(1-2):81-4. https://doi.org/10.1016/0304-3940(94)90361-1
  28. Jeon YT, Seo KS, Ro YJ et al. The role of nitric oxide synthase isoforms in neuropathic pain induced by nerve injury in rats. Korean J Anesthesiol. 2005;48(1):76-84. https://doi.org/10.4097/kjae.2005.48.1.76
  29. Hiura A. Is thermal nociception only sensed by the capsaicin receptor, TRPV1? Anat Sci Int. 2009;84(3):122-8. https://doi.org/10.1007/s12565-009-0048-8
  30. Caterina MJ, Leffler A, Malmberg AB et al. Impaired nociception and pain sensation in mice lacking the capsaicin receptor. Science. 2000;288(5464):306-13. https://doi.org/10.1126/science.288.5464.306
  31. Planells-Cases R, Garcia-Sanz N, Morenilla-Palao C et al. Functional aspects and mechanisms of TRPV1 involvement in neurogenic inflammation that leads to thermal hyperalgesia. Pflugers Arch. 2005;451(1):151-9. https://doi.org/10.1007/s00424-005-1423-5
  32. Doly S, Fischer J, Salio C et al. The vanilloid receptor-1 is expressed in rat spinal dorsal horn astrocytes. Neurosci Lett. 2004;357(2):123-6. https://doi.org/10.1016/j.neulet.2003.12.051
  33. Birder LA, Kanai AJ, de Groat WC et al. Vanilloid receptor expression suggests a sensory role for urinary bladder epithelial cells. Proc Natl Acad Sci U S A. 2001;98(23):13396- 401. https://doi.org/10.1073/pnas.231243698
  34. Sanchez JF, Krause JE, Cortright DN. The distribution and regulation of vanilloid receptor VR1 and VR1 5′ splice variant RNA expression in rat. Neuroscience. 2001;107(3):373-81. https://doi.org/10.1016/S0306-4522(01)00373-6