RESUMO
Despite many decades of drug development, effective therapies for neuropathic pain remain elusive. The recent recognition of spinal cord glia and glial pro-inflammatory cytokines as important contributors to neuropathic pain suggests an alternative therapeutic strategy; that is, targeting glial activation or its downstream consequences. While several glial-selective drugs have been successful in controlling neuropathic pain in animal models, none are optimal for human use. Thus the aim of the present studies was to explore a novel approach for controlling neuropathic pain. Here, an adeno-associated viral (serotype II; AAV2) vector was created that encodes the anti-inflammatory cytokine, interleukin-10 (IL-10). This anti-inflammatory cytokine is known to suppress the production of pro-inflammatory cytokines. Upon intrathecal administration, this novel AAV2-IL-10 vector was successful in transiently preventing and reversing neuropathic pain. Intrathecal administration of an AAV2 vector encoding beta-galactosidase revealed that AAV2 preferentially infects meningeal cells surrounding the CSF space. Taken together, these data provide initial support that intrathecal gene therapy to drive the production of IL-10 may prove to be an efficacious treatment for neuropathic pain.
Assuntos
Dependovirus/genética , Terapia Genética/métodos , Mediadores da Inflamação/fisiologia , Interleucina-10/biossíntese , Interleucina-10/genética , Nervo Isquiático/fisiopatologia , Ciática/prevenção & controle , Ciática/fisiopatologia , Animais , Dependovirus/fisiologia , Modelos Animais de Doenças , Vetores Genéticos/administração & dosagem , Vetores Genéticos/uso terapêutico , Humanos , Inflamação/metabolismo , Inflamação/prevenção & controle , Inflamação/virologia , Injeções Espinhais , Interleucina-10/fisiologia , Masculino , Ratos , Ratos Sprague-Dawley , Ciática/metabolismoRESUMO
UNLABELLED: Glia are now recognized as important contributors in pathological pain creation and maintenance. Spinal cord glia exhibit extensive gap junctional connectivity, raising the possibility that glia are involved in the contralateral spread of excitation resulting in mirror image pain. In the present experiments, the gap junction decoupler carbenoxolone was administered intrathecally after induction of neuropathic pain in response to sciatic nerve inflammation (sciatic inflammatory neuropathy) or partial nerve injury (chronic constriction injury). In both neuropathic pain models, a low dose of carbenoxolone reversed mirror image mechanical allodynia, while leaving ipsilateral mechanical allodynia unaffected. Ipsilateral thermal hyperalgesia was briefly attenuated. Critically, blockade of mechanical allodynia and thermal hyperalgesia was not observed in response to intrathecal glycyrrhizic acid, a compound similar to carbenoxolone in all respects but it does not decouple gap junctions. Thus, blockade of mechanical allodynia and thermal hyperalgesia by carbenoxolone does appear to reflect an effect on gap junctions. Examination of carbenoxolone's effects on intrathecal human immunodeficiency virus type 1 gp120 showed that blockade of pain facilitation might result, at least in part, via suppression of interleukin-1 and, in turn, interleukin-6. These data provide the first suggestion that spread of excitation via gap junctions might contribute importantly to inflammatory and traumatic neuropathic pain. PERSPECTIVE: The current studies provide evidence for involvement of gap junctions in spinal cord pain facilitation. Intrathecal carbenoxolone, a gap junction decoupler, reversed neuropathy-induced mirror image pain and intrathecal gp120-induced allodynia. In addition, it decreased gp120-induced proinflammatory cytokines. This suggests gap junction activation might lead to proinflammatory cytokine release by distantly activated glia.
Assuntos
Junções Comunicantes/fisiologia , Dor/fisiopatologia , Células do Corno Posterior/fisiologia , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Antiulcerosos/farmacologia , Carbenoxolona/farmacologia , Doença Crônica , Junções Comunicantes/efeitos dos fármacos , Ácido Glicirrízico/farmacologia , Proteína gp120 do Envelope de HIV/farmacologia , Hiperalgesia/fisiopatologia , Injeções Espinhais , Interleucina-1/metabolismo , Masculino , Síndromes de Compressão Nervosa/fisiopatologia , Ratos , Ratos Sprague-Dawley , Neuropatia Ciática/fisiopatologia , Organismos Livres de Patógenos EspecíficosRESUMO
It has become clear that spinal cord glia (microglia and astrocytes) importantly contribute to the creation of exaggerated pain responses. One model used to study this is peri-spinal (intrathecal, i.t.) administration of gp120, an envelope protein of HIV-1 known to activate glia. Previous studies demonstrated that i.t. gp120 produces pain facilitation via the release of glial proinflammatory cytokines. The present series of studies tested whether spinal nitric oxide (NO) contributes to i.t. gp120-induced mechanical allodynia and, if so, what effect NO has on spinal proinflammatory cytokines. gp120 stimulation of acutely isolated lumbar dorsal spinal cords released NO as well as proinflammatory cytokines (tumor necrosis factor-alpha, interleukin-1beta (IL1), interleukin-6 (IL6)), thus identifying NO as a candidate mediator of gp120-induced behavioral effects. Behaviorally, identical effects were observed when gp120-induced mechanical allodynia was challenged by i.t. pre-treatment with either a broad-spectrum nitric oxide synthase (NOS) inhibitor (L-NAME) or 7-NINA, a selective inhibitor of NOS type-I (nNOS). Both abolished gp120-induced mechanical allodynia. While the literature pre-dominantly documents that proinflammatory cytokines stimulate the production of NO rather than the reverse, here we show that gp120-induced NO increases proinflammatory cytokine mRNA levels (RT-PCR) and both protein expression and protein release (serial ELISA). Furthermore, gp120 increases mRNA for IL1 converting enzyme and matrix metalloproteinase-9, enzymes responsible for activation and release of proinflammatory cytokines.
Assuntos
Citocinas/fisiologia , Proteína gp120 do Envelope de HIV/farmacologia , Mediadores da Inflamação/fisiologia , Óxido Nítrico Sintase/metabolismo , Dor/enzimologia , Animais , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/fisiologia , Masculino , Óxido Nítrico Sintase/fisiologia , Óxido Nítrico Sintase Tipo I , Dor/virologia , Ratos , Ratos Sprague-DawleyRESUMO
While chronic use of indwelling micromachined neural prosthetic devices has great potential, the development of reactive responses around them results in a decrease in electrode function over time. Since the cellular events responsible for these responses may be anti-inflammatory in nature, we have tested the effectiveness of dexamethasone and cyclosporin A as potential drugs for developing intervention strategies following insertion of single-shank micromachined silicon devices. Peripheral injection of dexamethasone was effective in attenuating increased expression of glial fibrillary acidic protein and astrocyte hyperplasia observed during both initial- and sustained-reactive responses observed at one and six weeks post insertion, respectively. Peripheral injection of cyclosporin A had no positive effect. If anything, application of this drug increased the early reactive response. Effectiveness of local release of dexamethasone in rat neocortex was tested by inserting ribbons of poly (ethyl-vinyl) acetate containing 35% (w/w) dexamethasone. Initial concentrations of dexamethasone were similar to those obtained by peripheral injection. Local drug release provided continued control of cellular reactive responses during the six-week study period. These results demonstrate that peripheral delivery of dexamethasone can be used to control reactive responses and that local drug delivery by slow-release from biocompatible polymers may be a more effective method of drug intervention. Incorporating these strategies on micromachined devices may provide an intervention strategy that will insure the chronic functioning of electrodes on intracortical neuroprosthetic devices.
Assuntos
Materiais Revestidos Biocompatíveis/uso terapêutico , Ciclosporina/administração & dosagem , Dexametasona/administração & dosagem , Eletrodos/efeitos adversos , Próteses e Implantes/efeitos adversos , Infecções Relacionadas à Prótese/tratamento farmacológico , Animais , Preparações de Ação Retardada/administração & dosagem , Injeções Subcutâneas , Masculino , Doenças do Sistema Nervoso/reabilitação , Infecções Relacionadas à Prótese/prevenção & controle , RatosRESUMO
Previous studies have found inconsistent relationships between restrained eating, dieting, and cortisol. The present study was designed to clarify the relationship between self-reported restrained eating and cortisol using multiple measures of dietary restraint. Eighty-five college-age women completed the Restraint Scale (RS) and the Cognitive Restraint Scale of the Three Factor Eating Questionnaire (TFEQ-R) and provided a saliva sample for analysis of cortisol. Both measures of restraint were positively associated with elevated levels of salivary cortisol, although the TFEQ-R was more strongly associated than the RS. Restrained eating, characterized by largely unsuccessful efforts to control eating, may lead to elevated cortisol levels.