J Korean Med Sci.  2015 Aug;30(8):1189-1196. 10.3346/jkms.2015.30.8.1189.

Mechanical Antiallodynic Effect of Intrathecal Nefopam in a Rat Neuropathic Pain Model

Affiliations
  • 1Department of Anesthesia and Pain Medicine, Pusan National University School of Medicine; Research Institute for Convergence of biomedical science and technology Pusan National University Yangsan Hospital, Yangsan, Korea. byeongj@pusan.ac.kr
  • 2Division of Meridian and Structural Medicine, Pusan National University School of Korean Medicine, Yangsan, Korea.

Abstract

Nefopam has a pharmacologic profile distinct from that of opioids or other anti-inflammatory drugs. Several recent studies demonstrate that nefopam has a mechanism of action similar to those of anti-depressants and anticonvulsants for treating neuropathic pain. The present study investigates the mechanical antiallodynic effect of nefopam using immunohistochemical study and western blot analysis in a rat neuropathic pain model. Twenty-eight male Sprague-Dawley rats were subjected to left fifth lumbar (L5) spinal nerve ligation and intrathecal catheter implantation, procedures which were not performed on the 7 male Sprague-Dawley rats in the sham surgery group (group S). Nefopam, either 10 or 100 microg/kg (group N10 or N100, respectively), and normal saline (group C) were intrathecally administered into the catheter every day for 14 days. The mechanical allodynic threshold of intrathecal nefopam was measured using a dynamic plantar aesthesiometer. Immunohistochemistry targeting cluster of differentiation molecule 11b (CD11b) and glial fibrillary acidic protein (GFAP) was performed on the harvested spinal cord at the level of L5. Extracellular signal-regulated kinase 1/2 (ERK 1/2) and cyclic adenosine monophosphate response element binding protein (CREB) were measured using western blot analysis. The N10 and N100 groups showed improved mechanical allodynic threshold, reduced CD11b and GFAP expression, and attenuated ERK 1/2 and CREB in the affected L5 spinal cord. In conclusion, intrathecal nefopam reduced mechanical allodynia in a rat neuropathic pain model. Its mechanical antiallodynic effect is associated with inhibition of glial activation and suppression of the transcription factors' mitogen-activated protein kinases in the spinal cord.

Keyword

Mechanical Allodynia; Nefopam; Neuropathic Pain; Spinal Nerve Ligation

MeSH Terms

Analgesics, Non-Narcotic/administration & dosage
Animals
Dose-Response Relationship, Drug
Hyperalgesia/*drug therapy/etiology/*physiopathology
Injections, Spinal
Male
Nefopam/*administration & dosage
Neuralgia/complications/*drug therapy/*physiopathology
Pain Measurement/drug effects
Pain Perception/*drug effects
Rats
Rats, Sprague-Dawley
Treatment Outcome
Analgesics, Non-Narcotic
Nefopam

Figure

  • Fig. 1 Behavioral assessments. (A) Schematic representation of behavioral assessments after intrathecal injection of nefopam. (B) Time course change of hind limb mechanical sensitivity to a dynamic plantar aesthesiometer after intrathecal injection of nefopam. (C) The change in withdrawal threshold following intrathecal injection of nefopam (10 or 100 µg/kg) every day or normal saline in rats receiving spinal nerve ligation (SNL). *P < 0.05 compared to control by one-way analysis of variance (ANOVA); †P < 0.05 compared to baseline by repeated measure ANOVA. S: sham surgery group, C: normal saline group, N10: 10 µg/kg nefopam group, N100: 100 µg/kg nefopam group, Initial: before SNL, Post-SNL: 3rd day after SNL, Post-ICI: 1st day after surgical intrathecal catheter insertion, Day-7: 7th day after intrathecal nefopam injection, Day-14: 14th day after intrathecal nefopam injection.

  • Fig. 2 Photographs (40 × magnification) illustrating the effect of nefopam on spinal immunoreactivity to cluster of differentiation molecule 11b (CD11b) after left L5 spinal nerve ligation. The immunohistochemical intensity of CD11b in the L5 segment decreased in a dose dependent manner following nefopam administration. (A) sham surgery group, (B) normal saline group, (C) 10 µg/kg of nefopam group, (D) 100 µg/kg of nefopam group.

  • Fig. 3 Photographs (40 × magnification) illustrating the effect of nefopam on spinal immunoreactivity to glial fibrillary acidic protein (GFAP) after left L5 spinal nerve ligation. The immunohistochemical intensity of GFAP in the L5 segment decreased in a dose dependent manner following nefopam. (A) sham surgery group, (B) normal saline group, (C) 10 µg/kg nefopam group, (D) 100 µg/kg nefopam group.

  • Fig. 4 Changes in the relative mean optical density (MOD) ratio for (A) CD11b and (B) GFAP after left L5 spinal nerve ligation. The relative MOD ratio increased in the spinal cord side ipsilateral to the spinal nerve ligation in rats. The relative MOD ratio decreased in a dose-dependent manner in rats administered intrathecal nefopam (N10 and N100) compared to the ratio in rats administered intrathecal saline (group C). *P < 0.05 compared to the S group; †P < 0.05 compared to the C group. S: sham surgery group, C: normal saline group, N10: 10 µg/kg nefopam group, N100: 100 µg/kg nefopam group.

  • Fig. 5 The expression of extracellular signal-regulated kinase 1/2 (ERK 1/2) and cyclic adenosine monophosphate response element-binding (CREB) measured by immunoblotting. (A) Representative immunoblot images for ERK 1/2 and CREB protein in spinal cord from the S, C, N10, and N100 groups. (B, C) Densitometric quantifications of ERK 1/2 and CREB band intensities. *P < 0.05 compared to the S group; †P < 0.05 compared to the C group. S: sham surgery group, C: normal saline group, N10: 10 µg/kg nefopam group, N100: 100 µg/kg nefopam group.


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Reference

1. Lee HJ, Shin SW, Jang HJ. The combined antiallodynic effect of gabapentin and milnacipran in a rat neuropathic pain model. Korean J Pain. 2007; 20:8–14.
2. Sindrup SH, Jensen TS. Efficacy of pharmacological treatments of neuropathic pain: an update and effect related to mechanism of drug action. Pain. 1999; 83:389–400.
3. Hunter JC, Gogas KR, Hedley LR, Jacobson LO, Kassotakis L, Thompson J, Fontana DJ. The effect of novel anti-epileptic drugs in rat experimental models of acute and chronic pain. Eur J Pharmacol. 1997; 324:153–160.
4. Idänpään-Heikkilä JJ, Guilbaud G. Pharmacological studies on a rat model of trigeminal neuropathic pain: baclofen, but not carbamazepine, morphine or tricyclic antidepressants, attenuates the allodynia-like behaviour. Pain. 1999; 79:281–290.
5. Iyengar S, Webster AA, Hemrick-Luecke SK, Xu JY, Simmons RM. Efficacy of duloxetine, a potent and balanced serotonin-norepinephrine reuptake inhibitor in persistent pain models in rats. J Pharmacol Exp Ther. 2004; 311:576–584.
6. Guirimand F, Dupont X, Bouhassira D, Brasseur L, Chauvin M. Nefopam strongly depresses the nociceptive flexion (R(III)) reflex in humans. Pain. 1999; 80:399–404.
7. Biella GE, Groppetti A, Novelli A, Fernández-Sánchez MT, Manfredi B, Sotgiu ML. Neuronal sensitization and its behavioral correlates in a rat model of neuropathy are prevented by a cyclic analog of orphenadrine. J Neurotrauma. 2003; 20:593–601.
8. Gregori-Puigjané E, Setola V, Hert J, Crews BA, Irwin JJ, Lounkine E, Marnett L, Roth BL, Shoichet BK. Identifying mechanism-of-action targets for drugs and probes. Proc Natl Acad Sci U S A. 2012; 109:11178–11183.
9. Kim SH, Chung JM. An experimental model for peripheral neuropathy produced by segmental spinal nerve ligation in the rat. Pain. 1992; 50:355–363.
10. Zhuang ZY, Gerner P, Woolf CJ, Ji RR. ERK is sequentially activated in neurons, microglia, and astrocytes by spinal nerve ligation and contributes to mechanical allodynia in this neuropathic pain model. Pain. 2005; 114:149–159.
11. Song XS, Cao JL, Xu YB, He JH, Zhang LC, Zeng YM. Activation of ERK/CREB pathway in spinal cord contributes to chronic constrictive injury-induced neuropathic pain in rats. Acta Pharmacol Sin. 2005; 26:789–798.
12. Ma W, Quirion R. Increased phosphorylation of cyclic AMP response element-binding protein (CREB) in the superficial dorsal horn neurons following partial sciatic nerve ligation. Pain. 2001; 93:295–301.
13. Kang G, Choi KY, Lee MS, Ahn YJ, Kang SS, Cheong IY, Chun W, Kim SS. Minocycline attenuates the development of allodynia: an immunohistochemical study on CD11b, GFAP and c-Fos in the spinal dorsal horn in SD rat. Korean J Pathol. 2004; 38:311–318.
14. Saghaei E, Moini Zanjani T, Sabetkasaei M, Naseri K. Enhancement of antinociception by co-administrations of nefopam, morphine, and nimesulide in a rat model of neuropathic pain. Korean J Pain. 2012; 25:7–15.
15. Fuller RW, Snoddy HD. Evaluation of nefopam as a monoamine uptake inhibitor in vivo in mice. Neuropharmacology. 1993; 32:995–999.
16. Rosland JH, Hole K. The effect of nefopam and its enantiomers on the uptake of 5-hydroxytryptamine, noradrenaline and dopamine in crude rat brain synaptosomal preparations. J Pharm Pharmacol. 1990; 42:437–438.
17. Verleye M, André N, Heulard I, Gillardin JM. Nefopam blocks voltage-sensitive sodium channels and modulates glutamatergic transmission in rodents. Brain Res. 2004; 1013:249–255.
18. Ren K, Dubner R. Descending modulation in persistent pain: an update. Pain. 2002; 100:1–6.
19. Millan MJ. Descending control of pain. Prog Neurobiol. 2002; 66:355–474.
20. Fasmer OB, Berge OG, Jørgensen HA, Hole K. Antinociceptive effects of (+/-)-, (+)- and (-)-nefopam in mice. J Pharm Pharmacol. 1987; 39:508–511.
21. Piercey MF, Schroeder LA. Spinal and supraspinal sites for morphine and nefopam analgesia in the mouse. Eur J Pharmacol. 1981; 74:135–140.
22. Cho SY, Park AR, Yoon MH, Lee HG, Kim WM, Choi JI. Antinociceptive effect of intrathecal nefopam and interaction with morphine in formalin-induced pain of rats. Korean J Pain. 2013; 26:14–20.
23. Marchand F, Perretti M, McMahon SB. Role of the immune system in chronic pain. Nat Rev Neurosci. 2005; 6:521–532.
24. Mollace V, Colasanti M, Muscoli C, Lauro GM, Iannone M, Rotiroti D, Nistico G. The effect of nitric oxide on cytokine-induced release of PGE2 by human cultured astroglial cells. Br J Pharmacol. 1998; 124:742–746.
25. Kreutzberg GW. Microglia: a sensor for pathological events in the CNS. Trends Neurosci. 1996; 19:312–318.
26. Raghavendra V, Tanga F, Rutkowski MD, DeLeo JA. Anti-hyperalgesic and morphine-sparing actions of propentofylline following peripheral nerve injury in rats: mechanistic implications of spinal glia and proinflammatory cytokines. Pain. 2003; 104:655–664.
27. Ledeboer A, Sloane EM, Milligan ED, Frank MG, Mahony JH, Maier SF, Watkins LR. Minocycline attenuates mechanical allodynia and proinflammatory cytokine expression in rat models of pain facilitation. Pain. 2005; 115:71–83.
28. Crown ED, Ye Z, Johnson KM, Xu GY, McAdoo DJ, Hulsebosch CE. Increases in the activated forms of ERK 1/2, p38 MAPK, and CREB are correlated with the expression of at-level mechanical allodynia following spinal cord injury. Exp Neurol. 2006; 199:397–407.
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