Supraspinal kynurenine pathway contributes to the maintenance of neuropathic pain

Author(s)

Dênis Augusto Santana Reis

• Document type: Master's Dissertation
• Press: Ribeirão Preto.
• Institution: Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC)
• Defense date: 2015-02-03

Examining board members

Thiago Mattar Cunha; Joice Maria da Cunha; Helio Zangrossi Junior

• Advisor: Thiago Mattar Cunha

Abstract

Introduction: One factor that may contribute to the development of neuropathic pain is the negative modulation of the descending pain pathway by increased degradation of the activation of tryptophan by enzyme indoleamine 2,3-dioxygenase1 (IDO1) or activation of the descending facilitatory pain pathway for a glutamate agonist produced by the enzyme kynurenine 3 monooxygenase (KMO). Aim: We evaluate the role of IDO1 and KMO in the periaqueductal gray (PAG) and the rostral ventromedial medulla (RVM) in the development of neuropathic pain in mice induced by SNI model. Methods: Induction of experimental neuropathy was performed according to (Bourquin et al. 2006). The expression of IDO1 and KMO was carried out by Western blotting technique. The drug administration was performed orally, intraperitoneally and intracerebroventricularly (i.c.v) Results. We observed increased IDO1 expression in the RVM (7 days) and PAG (3, 7, 14 and 21 days) after SNI. The microinjection Norharmane in i.c.v. space reduced mechanical hypersensitivity in the 7, 14 and 21 days after SNI. Corroborating these findings, mice deficient for the enzyme IDO1 undergoing SNI did not develop mechanical hypersensitivity. Furthermore, the KMO expression was significantly increased in the 7 and 14 days in the RVM and 7 days in PAG after SNI. Therefore, oral administration of JM6, prodrug slow release from Ro61-8048 or Ro61-8048 (KMO inhibitors) within i.c.v. significantly reduced the mechanical hypersensitivity at day 7, 14 or 21 after SNI. Knowing that the expression of IDO1 enzyme is modulated by IFN- cytokine, it was found that animals deficient for IFN- cytokine have reduced mechanical hypersensitivity. Moreover, IFN- ko animals have reduced expression of IDO1 RVM 7 days and 14 days after SNI in the PAG. In addition, microinjection of increasing doses of IFN- in i.c.v. induced mechanical hyperalgesia. We also found that CD4 + KO animals, but not CD8 + KO animals showed reduced expression of the enzyme IDO1 RVM and PAG and consequently lower mechanical hypersensitivity after SNI. The microinjection of the main metabolites of kynurenine pathway into the i.c.v. spaces induced mechanical hypersensitivity, QUIN being the most potent. We suggest that the activation of the kynurenine pathway was dependent of NMDA receptor activation, whereas the spot pre-treatment with MK801 (selective NMDA receptor antagonist) reverses the effects induced by noxious metabolites. After that, the microinjection into i.c.v. spaces of MK801 reduced mechanical hypersensitivity after SNI. Furthermore, nociceptive effect induced by QUIN depends activation of the descending facilitatory. We found that the neuropathic animals exhibit depressive-like behavior and this behavior is not observed in IFN- KO and CD4KO mice. Finally, we evaluate the participation of kynurenine pathway in the development of depressive-like behavior associated with SNI and found that this behavior depends on the activation of IDO1 and KMO Conclusion: These results suggest that IDO1 and KMO enzyme, located in supraspinal regions play a role in the development of neuropathic pain as well as comorbidity depression. Furthermore, the expression of IDO1 are dependent on signaling via cytokine IFN- and CD4+ cells. The mechanism responsible for the development of neuropathic hypersensitivity is due to both reduced levels of tryptophan/5-HT decrease the descending inhibitory pain pathway efficiency, as the increased levels of QUIN, which activates the descending facilitatory pain pathway. (AU)

• FAPESP's process: 13/07968-0 - The role of indoleamine 2,3-dioxygenase in supraspinal structures in the induction and maintenance of neuropathic pain
• Grantee: Dênis Augusto Santana Reis
• Support Opportunities: Scholarships in Brazil - Master