RESUMO
Peripheral nerve injury sensitizes a complex network of spinal cord dorsal horn (DH) neurons to produce allodynia and neuropathic pain. The identification of a druggable target within this network has remained elusive, but a promising candidate is the neuropeptide Y (NPY) Y1 receptor-expressing interneuron (Y1-IN) population. We report that spared nerve injury (SNI) enhanced the excitability of Y1-INs and elicited allodynia (mechanical and cold hypersensitivity) and affective pain. Similarly, chemogenetic or optogenetic activation of Y1-INs in uninjured mice elicited behavioral signs of spontaneous, allodynic, and affective pain. SNI-induced allodynia was reduced by chemogenetic inhibition of Y1-INs, or intrathecal administration of a Y1-selective agonist. Conditional deletion of Npy1r in DH neurons, but not peripheral afferent neurons prevented the anti-hyperalgesic effects of the intrathecal Y1 agonist. We conclude that spinal Y1-INs are necessary and sufficient for the behavioral symptoms of neuropathic pain and represent a promising target for future pharmacotherapeutic development of Y1 agonists.
Assuntos
Hiperalgesia , Neuralgia , Camundongos , Animais , Hiperalgesia/tratamento farmacológico , Neuropeptídeo Y/genética , Neuropeptídeo Y/farmacologia , Neuralgia/tratamento farmacológico , Neurônios , Medula EspinalRESUMO
COQ7 encodes a hydroxylase responsible for the penultimate step of coenzyme Q10 (CoQ10) biosynthesis in mitochondria. CoQ10 is essential for multiple cellular functions, including mitochondrial oxidative phosphorylation, lipid metabolism, and reactive oxygen species homeostasis. Mutations in COQ7 have been previously associated with primary CoQ10 deficiency, a clinically heterogeneous multisystemic mitochondrial disorder. We identified COQ7 biallelic variants in nine families diagnosed with distal hereditary motor neuropathy with upper neuron involvement, expending the clinical phenotype associated with defects in this gene. A recurrent p.Met1? change was identified in five families from Brazil with evidence of a founder effect. Fibroblasts isolated from patients revealed a substantial depletion of COQ7 protein levels, indicating protein instability leading to loss of enzyme function. High-performance liquid chromatography assay showed that fibroblasts from patients had reduced levels of CoQ10, and abnormal accumulation of the biosynthetic precursor DMQ10. Accordingly, fibroblasts from patients displayed significantly decreased oxygen consumption rates in patients, suggesting mitochondrial respiration deficiency. Induced pluripotent stem cell-derived motor neurons from patient fibroblasts showed significantly increased levels of extracellular neurofilament light protein, indicating axonal degeneration. Our findings indicate a molecular pathway involving CoQ10 biosynthesis deficiency and mitochondrial dysfunction in patients with distal hereditary motor neuropathy. Further studies will be important to evaluate the potential benefits of CoQ10 supplementation in the clinical outcome of the disease.
Assuntos
Doenças Mitocondriais , Humanos , Mitocôndrias/genética , Mitocôndrias/metabolismo , Doenças Mitocondriais/metabolismo , Neurônios Motores/metabolismo , Mutação/genética , Ubiquinona/genéticaRESUMO
Disability in leprosy is a direct consequence of damage to the peripheral nervous system which is usually worse in patients with no skin manifestations, an underdiagnosed subtype of leprosy known as primary neural leprosy. We evaluated clinical, neurophysiological and laboratory findings of 164 patients with definite and probable primary neural leprosy diagnoses. To better understand the disease progression and to improve primary neural leprosy clinical recognition we compared the characteristics of patients with short (≤12 months) and long (>12 months) disease duration. Positive and negative symptoms mediated by small-fibres were frequent at presentation (â¼95%), and symptoms tend to manifest first in the upper limbs (â¼68%). There is a consistent phenotypic variability between the aforementioned groups. Deep sensory modalities were spared in patients evaluated within the first 12 months of the disease, and were only affected in patients with longer disease duration (â¼12%). Deep tendon reflexes abnormalities were most frequent in patients with longer disease duration (P < 0.001), as well as motor deficits (P = 0.002). Damage to large fibres (sensory and motor) is a latter event in primary neural leprosy. Grade-2 disability and nerve thickening was also more frequent in cases with long disease duration (P < 0.001). Primary neural leprosy progresses over time and there is a marked difference in clinical phenotype between patients with short and long disease duration. Patients assessed within the first 12 months of symptom onset had a non-length-dependent predominant small-fibre sensory neuropathy, whilst patients with chronic disease presented an asymmetrical all diameter sensory-motor neuropathy and patchily decreased/absent deep tendon reflexes.
Assuntos
Hanseníase Tuberculoide , Hanseníase , Doenças do Sistema Nervoso Periférico , Humanos , Hanseníase/complicações , Hanseníase/diagnóstico , Hanseníase/patologia , Hanseníase Tuberculoide/diagnóstico , Hanseníase Tuberculoide/patologia , Doenças do Sistema Nervoso Periférico/diagnósticoRESUMO
Painful diabetic neuropathy (PDN) is a devastating neurological complication of diabetes. Methylglyoxal (MG) is a reactive metabolite whose elevation in the plasma corresponds to PDN in patients and pain-like behavior in rodent models of type 1 and type 2 diabetes. Here, we addressed the MG-related spinal mechanisms of PDN in type 2 diabetes using db/db mice, an established model of type 2 diabetes, and intrathecal injection of MG in conventional C57BL/6J mice. Administration of either a MG scavenger (GERP10) or a vector overexpressing glyoxalase 1, the catabolic enzyme for MG, attenuated heat hypersensitivity in db/db mice. In C57BL/6J mice, intrathecal administration of MG produced signs of both evoked (heat and mechanical hypersensitivity) and affective (conditioned place avoidance) pain. MG-induced Ca2+ mobilization in lamina II dorsal horn neurons of C57BL/6J mice was exacerbated in db/db, suggestive of MG-evoked central sensitization. Pharmacological and/or genetic inhibition of transient receptor potential ankyrin subtype 1 (TRPA1), adenylyl cyclase type 1 (AC1), protein kinase A (PKA), or exchange protein directly activated by cyclic adenosine monophosphate (Epac) blocked MG-evoked hypersensitivity in C57BL/6J mice. Similarly, intrathecal administration of GERP10, or inhibitors of TRPA1 (HC030031), AC1 (NB001), or Epac (HJC-0197) attenuated hypersensitivity in db/db mice. We conclude that MG and sensitization of a spinal TRPA1-AC1-Epac signaling cascade facilitate PDN in db/db mice. Our results warrant clinical investigation of MG scavengers, glyoxalase inducers, and spinally-directed pharmacological inhibitors of a MG-TRPA1-AC1-Epac pathway for the treatment of PDN in type 2 diabetes.
Assuntos
Adenilil Ciclases/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Neuropatias Diabéticas/metabolismo , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Aldeído Pirúvico/metabolismo , Canal de Cátion TRPA1/metabolismo , Animais , Aprendizagem da Esquiva/efeitos dos fármacos , Aprendizagem da Esquiva/fisiologia , Comportamento Animal/efeitos dos fármacos , Comportamento Animal/fisiologia , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Diabetes Mellitus Tipo 2/complicações , Masculino , Camundongos , Medição da Dor , Células do Corno Posterior/efeitos dos fármacos , Células do Corno Posterior/metabolismo , Aldeído Pirúvico/farmacologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologiaRESUMO
Tissue injury creates a delicate balance between latent pain sensitization (LS) and compensatory endogenous analgesia. Inhibitory G-protein-coupled receptor (GPCR) interactions that oppose LS, including µ-opioid receptor (MOR) or neuropeptide Y Y1 receptor (Y1R) activity, persist in the spinal cord dorsal horn (DH) for months, even after the resolution of normal pain thresholds. Here, we demonstrate that following recovery from surgical incision, a potent endogenous analgesic synergy between MOR and Y1R activity persists within DH interneurons to reduce the intensity and duration of latent postoperative hypersensitivity and ongoing pain. Failure of such endogenous GPCR signaling to maintain LS in remission may underlie the transition from acute to chronic pain states.
RESUMO
Ceramic honeycomb monoliths were washcoated with cryptomelane-type manganese oxides and their catalytic performance was evaluated in the oxidation of ethyl acetate. The effect of a mixture of ethyl acetate with toluene and of the presence of water vapour was also assessed.Different coating parameters, namely size of catalyst particles, number of immersions in the washcoating solution, presence of an initial coating with alumina, calcination temperature of this coating, as well as the amount of binding agent and ethanol in the washcoating solution were studied and optimized based on the catalytic activity of the structured catalyst. Small particles are required for a correct impregnation; however, since the smallest particles are less active, an intermediate size achieved the best catalytic results. Increasing the number of immersions over 3 did not significantly increase the catalytic activity of the structured catalyst. The presence of an initial coating with alumina and a binding agent (colloidal alumina) in the washcoating solution was found essential to increase the activity, whereas increasing the calcination temperature after the initial alumina coating above 500°C decreased the activity of the catalyst. The presence of ethanol in the washcoating solution did not significantly improve the activity of the structured catalyst.The optimized structured catalyst presented high catalytic activity in the removal of ethyl acetate (90% conversion into CO2 at 256°C) and high stability during 100â h of reaction. The addition of toluene or water vapour in the feed gas did not significantly affect the activity of the coated monolith.
Assuntos
Compostos Orgânicos Voláteis , Catálise , Cerâmica , Compostos de Manganês , Oxirredução , ÓxidosRESUMO
Pharmacological agents directed to either opioid receptors or peroxisome proliferator-activated receptor gamma (PPARγ) at peripheral tissues reduce behavioral signs of persistent pain. Both receptors are expressed in muscle tissue, but the contribution of PPARγ activation to muscle pain and its modulation by opioid receptors remains unknown. To address this question, we first tested whether the endogenous PPARγ ligand 15d-PGJ2 would decrease mechanical hyperalgesia induced by carrageenan administration into the gastrocnemius muscle of rats. Next, we used receptor antagonists to determine whether the antihyperalgesic effect of 15-deoxyΔ-12,14-prostaglandin J2 (15d-PGJ2) was PPARγ- or opioid receptor-dependent. Three hours after carrageenan, muscle hyperalgesia was quantified with the Randall-Selitto test. 15d-PGJ2 prevented carrageenan-induced muscle hyperalgesia in a dose-dependent manner. The antihyperalgesic effect of 15d-PGJ2 was dose-dependently inhibited by either the PPARγ antagonist, 2-chloro-5-nitro-N-phenylbenzamide, or by the opioid receptor antagonist, naloxone. We conclude that 15d-PGJ2 targets PPARγ and opioid receptors to prevent muscle hyperalgesia. We suggest that local PPARγ receptors are important pharmacological targets for inflammatory muscle pain.
Assuntos
Hiperalgesia/metabolismo , Fatores Imunológicos/farmacologia , Músculo Esquelético/efeitos dos fármacos , Mialgia/metabolismo , PPAR gama/efeitos dos fármacos , Prostaglandina D2/análogos & derivados , Anilidas/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Carragenina/toxicidade , Hiperalgesia/induzido quimicamente , Músculo Esquelético/metabolismo , Mialgia/induzido quimicamente , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , PPAR gama/antagonistas & inibidores , Prostaglandina D2/farmacologia , RatosRESUMO
Cryptomelane-type manganese oxides prepared by a solvent-free method were evaluated as catalysts for the oxidation of ethyl acetate, ethanol and toluene. The original catalyst (K-OMS-2) presented high catalytic activity for ethyl acetate and ethanol oxidation, achieving 90% conversion into CO2 around 200°C for both pollutants. Toluene was much harder to oxidize, requiring a temperature near 270°C for the same conversion. The original catalyst was mechanically treated in a ball mill at different intensities, in order to decrease the particle size for subsequent impregnation onto structured supports, as small particle sizes are usually recommended. The catalytic activity of the materials decreases with the increase in the severity of this treatment, which is related to the decrease of the surface area of the catalysts, since the other properties (phase purity, thermal stability, surface oxygen, average oxidation state and reactivity of the oxygen species) are similar among the catalysts with different ball milling treatments. For comparison, a platinum-based catalyst (1%-Pt/Al2O3) was also tested, which exhibited a high activity for toluene, but much lower activities for the two other volatile organic compounds tested. A long-term experiment, using ethanol as model pollutant, showed that the cryptomelane catalyst was stable for more than 100â h.
Assuntos
Compostos Orgânicos Voláteis , Catálise , Oxirredução , Platina , ToluenoRESUMO
Peripheral inflammation produces a long-lasting latent sensitization of spinal nociceptive neurons, that is, masked by tonic inhibitory controls. We explored mechanisms of latent sensitization with an established four-step approach: (1) induction of inflammation; (2) allow pain hypersensitivity to resolve; (3) interrogate latent sensitization with a channel blocker, mutant mouse, or receptor antagonist; and (4) disrupt compensatory inhibition with a receptor antagonist so as to reinstate pain hypersensitivity. We found that the neuropeptide Y Y1 receptor antagonist BIBO3304 reinstated pain hypersensitivity, indicative of an unmasking of latent sensitization. BIBO3304-evoked reinstatement was not observed in AC1 knockout mice and was prevented with intrathecal co-administration of a pharmacological blocker to the N-methyl-D-aspartate receptor (NMDAR), adenylyl cyclase type 1 (AC1), protein kinase A (PKA), transient receptor potential cation channel A1 (TRPA1), channel V1 (TRPV1), or exchange protein activated by cAMP (Epac1 or Epac2). A PKA activator evoked both pain reinstatement and touch-evoked pERK expression in dorsal horn; the former was prevented with intrathecal co-administration of a TRPA1 or TRPV1 blocker. An Epac activator also evoked pain reinstatement and pERK expression. We conclude that PKA and Epac are sufficient to maintain long-lasting latent sensitization of dorsal horn neurons that is kept in remission by the NPY-Y1 receptor system. Furthermore, we have identified and characterized 2 novel molecular signaling pathways in the dorsal horn that drive latent sensitization in the setting of chronic inflammatory pain: NMDARâAC1âPKAâTRPA1/V1 and NMDARâAC1âEpac1/2. New treatments for chronic inflammatory pain might either increase endogenous NPY analgesia or inhibit AC1, PKA, or Epac.