Background Carbon monoxide (CO) synthesized by heme oxygenase 1 (HO-1) exerts antinociceptive effects during inflammation but its role during neuropathic pain remains unknown. Our objective is to investigate the exact contribution of CO derived from HO-1 in the modulation of neuropathic pain and the mechanisms implicated. Methodology/Principal Findings We evaluated the antiallodynic and antihyperalgesic effects of CO following sciatic nerve injury in wild type (WT) or inducible nitric oxide synthase knockout (NOS2-KO) mice using two carbon monoxide-releasing molecules (CORM-2 and CORM-3) and an HO-1 inducer (cobalt protoporphyrin IX, CoPP) daily administered from days 10 to 20 after injury. The effects of CORM-2 and CoPP on the expression of HO-1, heme oxygenase 2 (HO-2), neuronal nitric oxide synthase (NOS1) and NOS2 as well as a microglial marker (CD11b/c) were also assessed at day 20 after surgery in WT and NOS2-KO mice. In WT mice, the main neuropathic pain symptoms induced by nerve injury were significantly reduced in a time-dependent manner by treatment with CO-RMs or CoPP. Both CORM-2 and CoPP treatments increased HO-1 expression in WT mice, but only CoPP stimulated HO-1 in NOS2-KO animals. The increased expression of HO-2 induced by nerve injury in WT, but not in NOS2-KO mice, remains unaltered by CORM-2 or CoPP treatments. In contrast, the over-expression of CD11b/c, NOS1 and NOS2 induced by nerve injury in WT, but not in NOS2-KO mice, were significantly decreased by both CORM-2 and CoPP treatments. These data indicate that CO alleviates neuropathic pain through the reduction of spinal microglial activation and NOS1/NOS2 over-expression. Conclusions/Significance This study reports that an interaction between the CO and nitric oxide (NO) systems is taking place following sciatic nerve injury and reveals that increasing the exogenous (CO-RMs) or endogenous (CoPP) production of CO may represent a novel strategy for the treatment of neuropathic pain.
References
[1]
Watkins LR, Maier SF (2003) Glia: a novel drug discovery target for clinical pain. Nat Rev Drug Discov 2: 973–985.
[2]
La Buda CJ, Koblish M, Tuthill P, Dolle RE, Little PJ (2006) Antinociceptive activity of the selective iNOS inhibitor AR-C102222 in rodent models of inflammatory, neuropathic and post-operative pain. Eur J Pain 10: 505–512.
[3]
Meller ST, Pechman PS, Gebhart GF, Maves TJ (1992) Nitric oxide mediates the thermal hyperalgesia produced in a model of neuropathic pain in the rat. Neuroscience 50: 7–10.
[4]
Schmidtko A, Gao W, K？nig P, Heine S, Motterlini R, et al. (2008) cGMP produced by NO-sensitive guanylyl cyclase essentially contributes to inflammatory and neuropathic pain by using targets different from cGMP-dependent protein kinase I. J Neurosci. 28: 8568–8576.
[5]
De Alba J, Clayton NM, Collins SD, Colthup P, Chessell I, et al. (2006) GW274150, a novel and highly selective inhibitor of the inducible isoform of nitric oxide synthase (iNOS), shows analgesic effects in rat models of inflammatory and neuropathic pain. Pain 120: 170–181.
[6]
Hervera A, Negrete R, Leánez S, Martín-Campos JM, Pol O (2010) The spinal cord expression of neuronal and inducible nitric oxide synthases and their contribution in the maintenance of neuropathic pain in mice. PLoS One 5: e14321.
[7]
Levy D, Zochodne DW (1998) Local nitric oxide synthase activity in a model of neuropathic pain. Eur J Neurosci 10: 1846–1855.
[8]
Kuboyama K, Tsuda M, Tsutsui M, Toyohara Y, Tozaki-Saitoh H, et al. (2011) Reduced spinal microglial activation and neuropathic pain after nerve injury in mice lacking all three nitric oxide synthases. Mol Pain 7: 50.
[9]
Hervera A, Negrete R, Leánez S, Martín-Campos J, Pol O (2010) The role of nitric oxide in the local antiallodynic and antihyperalgesic effects and expression of delta-opioid and cannabinoid-2 receptors during neuropathic pain in mice. J Pharmacol Exp Ther 334: 887–896.
[10]
Naik AK, Tandan SK, Kumar D, Dudhgaonkar SP (2006) Nitric oxide and its modulators in chronic constriction injury-induced neuropathic pain in rats. Eur J Pharmacol 530: 59–69.
[11]
Fan W, Huang F, Wu Z, Zhu X, Li D, et al. (2011) Carbon monoxide: A gas that modulates nociception. J Neurosci Res 89: 802–807.
[12]
Rosa AO, Egea J, Lorrio S, Rojo AI, Cuadrado A, et al. (2008) Nrf2-mediated haeme oxygenase-1 up-regulation induced by cobalt protoporphyrin has antinociceptive effects against inflammatory pain in the formalin test in mice. Pain 137: 332–339.
[13]
Steiner AA, Branco LG, Cunha FQ, Ferreira SH (2000) Role of the haeme oxygenase/carbon monoxide pathway in mechanical nociceptor hypersensitivity. Br J Pharmacol 132: 1673–1682.
[14]
Li X, Clark JD (2000) The role of heme oxygenase in neuropathic and incisional pain. Anesth Analg 90: 677–682.
[15]
Li X, Clark JD (2003) Heme oxygenase type 2 participates in the development of chronic inflammatory and neuropathic pain. J Pain 4: 101–107.
[16]
Alcaraz MJ, Habib A, Lebret M, Créminon C, Lévy-Toledano S, et al. (2000) Enhanced expression of haem oxygenase-1 by nitric oxide and antiinflammatory drugs in NIH 3T3 fibroblasts. Br J Pharmacol 130: 57–64.
[17]
Foresti R, Clark JE, Green CJ, Motterlini R (1997) Thiol compounds interact with nitric oxide in regulating heme oxygenase-1 induction in endothelial cells: Involvement of superoxide and peroxynitrite anions. J Biol Chem 272: 18411–18417.
[18]
Kim YM, Bergonia HA, Müller C, Pitt BR, Watkins WD, et al. (1995) Loss and degradation of enzyme-bound heme induced by cellular nitric oxide synthesis. J Biol Chem 270: 5710–5713.
[19]
Lee SK, Park DY, Lee HJ, Lee J, Choi MK, et al. (2007) Functional interaction between nitric oxide-induced iron homeostasis and heme oxygenase-1 in immortalized and malignant oral keratinocytes. Cancer Lett 249: 283–293.
[20]
Clark JE, Naughton P, Shurey S, Green CJ, Johnson TR, et al. (2003) Cardioprotective actions by a water-soluble carbon monoxide-releasing molecule. Circ Res 93: e2–8.
[21]
Motterlini R, Clark JE, Foresti R, Sarathchandra P, Mann BE, et al. (2002) Carbon monoxide-releasing molecules: characterization of biochemical and vascular activitis. Circ Res 90: e17–24, 2002.
[22]
Motterlini R, Otterbein LE (2010) The therapeutic potential of carbon monoxide. Nat Rev Drug Discov 9: 728–743.
[23]
Sawle P, Foresti R, Mann BE, Johnson TR, Green CJ, et al. (2005) Carbon monoxide-releasing molecules (CO-RMs) attenuate the inflammatory response elicited by lipopolysaccharide in RAW264.7 murine macrophages. Br J Pharmacol 145: 800–810.
[24]
Guillén MI, Megías J, Clérigues V, Gomar F, Alcaraz MJ (2008) The CO-releasing molecule CORM-2 is a novel regulator of the inflammatory process in osteoarthritic chondrocytes. Rheumatology 47: 1323–1328.
[25]
Mika J, Osikowicz M, Rojewska E, Korostynski M, Wawrzczak-Bargiela A, et al. (2009) Differential activation of spinal microglial and astroglial cells in a mouse model of peripheral neuropathic pain. Eur J Pharmacol 623: 65–72.
[26]
Nascimento CG, Branco LG (2007) Role of the peripheral heme oxygenase-carbon monoxide pathway on the nociceptive response of rats to the formalin test: evidence for a cGMP signaling pathway. Eur J Pharmacol 556: 55–61.
[27]
Maicas N, Ferrándiz ML, Devesa I, Motterlini R, Koenders MI, et al. (2010) The CO-releasing molecule CORM-3 protects against articular degradation in the K/BxN serum transfer arthritis model. Eur J Pharmacol 634: 184–191.
[28]
Megías J, Guillén MI, Clérigues V, Rojo AI, Cuadrado A, et al. (2009) Heme oxygenase-1 induction modulates microsomal prostaglandin E synthase-1 expression and prostaglandin E(2) production in osteoarthritic chondrocytes. Biochem Pharmacol 77: 1806–1813.
[29]
Gordh T, Sharma HS, Azizi M, Alm P, Westman J (2000) Spinal nerve lesion induces upregulation of constitutive isoform of heme oxygenase in the spinal cord. An immunohistochemical investigation in the rat. Amino Acids 19: 373–381.
[30]
Watkins LR, Milligan ED, Maier SF (2001) Glial activation: a driving force for pathological pain. Trends Neurosci 24: 450–455.
[31]
Bani-Hani MG, Greenstein D, Mann BE, Green CJ, Motterlini R (2006) A carbon monoxide-releasing molecule (CORM-3) attenuates lipopolysaccharide- and interferon-gamma-induced inflammation in microglia. Pharmacol Rep 58: 132–144.
[32]
Wilkinson WJ, Kemp PJ (2011) The carbon monoxide donor, CORM-2, is an antagonist of ATP-gated, human P2X4 receptors. Purinergic Signal 7: 57–64.
[33]
Tsuda M, Shigemoto-Mogami Y, Koizumi S, Mizokoshi A, Kohsaka S, et al. (2003) P2X4 receptors induced in spinal microglia gate tactile allodynia after nerve injury. Nature 424: 778–783.
[34]
Bennett GJ, Xie YK (1988) A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man: Pain. 33: 87–107.
[35]
Chaplan SR, Bach FW, Pogrel JW, Chung JM, Yaksh TL (1994) Quantitative assessment of tactile allodynia in the rat paw. J Neurosci Methods 53: 55–63.
[36]
Hargreaves K, Dubner R, Brown F, Flores C, Joris J (1988) A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia. Pain 32: 77–88.
[37]
Devesa I, Ferrándiz ML, Terencio MC, Joosten LA, van den Berg WB, et al. (2005) Influence of heme oxygenase 1 modulation on the progression of murine collagen-induced arthritis. Arthritis Rheum 52: 3230–3238.
