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1.
Neurol Res ; 43(8): 683-692, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33866950

RESUMO

OBJECTIVE: Phospholipase A2 (PLA2) plays an important role in regulating the production of arachidonic acid and various eicosanoids. The aim of our study was to investigate the analgesic mechanisms of calcium-dependent cytosolic phospholipase A2 and calcium-independent PLA2 (iPLA2) inhibitors in the spinal cord in a rat model of neuropathic pain. METHODS: Lumbar 5 spinal nerve ligation was performed in male Sprague-Dawley rats to develop a peripheral neuropathic pain model. Paw withdrawal thresholds in response to von Frey filaments, brush, pressure, and pinch were measured. Lumbar wide dynamic range neuronal firing rates and iPLA2 subtype expression were measured by in vivo extracellular recording and double immunofluorescence staining, respectively. RESULTS: In our rat models, oral administration of prednisolone, a non-selective PLA2 inhibitor, and intrathecal injection of bromoenolactone, a iPLA2 inhibitor, significantly increased the ipsilateral hindpaw withdrawal thresholds in response to von Frey filament stimulation, but intrathecal injection of arachidonyl trifluoromethyl ketone, a selective cytosolic PLA2 inhibitor, did not show significant changes. In spinal dorsal horn neurons, bromoenolactone reduced neuronal firing rates in response to withdrawal stimulation and spontaneous firing rates in the ipsilateral side of the spinal dorsal horn. In addition, the expression of iPLA2 was co-localized with astrocytes and neurons on the ipsilateral side of the dorsal horn in rats that underwent spinal nerve ligation. DISCUSSION: These data suggest that selective iPLA2 inhibitor produce analgesia in neuropathic rats by reducing central sensitization in the dorsal horn.


Assuntos
Analgésicos/administração & dosagem , Inibidores Enzimáticos/administração & dosagem , Neuralgia/tratamento farmacológico , Fosfolipases A2 Independentes de Cálcio/antagonistas & inibidores , Corno Dorsal da Medula Espinal/efeitos dos fármacos , Administração Oral , Animais , Anti-Inflamatórios/administração & dosagem , Ácidos Araquidônicos/administração & dosagem , Injeções Espinhais , Masculino , Neuralgia/enzimologia , Fosfolipases A2 Independentes de Cálcio/metabolismo , Prednisolona/administração & dosagem , Ratos , Ratos Sprague-Dawley , Corno Dorsal da Medula Espinal/enzimologia
2.
Biomed Pharmacother ; 118: 109299, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31387001

RESUMO

We have recently demonstrated that the neurosteroid-metabolizing enzyme, cytochrome P450c17 is increased in spinal astrocytes contributing to the development of mechanical allodynia in chronic constriction injury (CCI)-induced neuropathic mice. However, the mechanisms by which spinal P450c17 modulates pathological changes in astrocytes remain unclear. In this study we investigated whether P450c17 modulates astrocyte activation and whether this process is mediated by spinal p38 mitogen-activated protein kinase phosphorylation ultimately leading to the development of mechanical allodynia in CCI mice. Sciatic nerve injury induced a significant increase in glial fibrillary acidic protein (GFAP) expression in the superficial dorsal horn (SDH, laminae I-II) and nucleus proprius (NP, laminae III-IV) regions of the spinal cord dorsal horn. Repeated daily (from days 0-3 post-surgery) intrathecal administration of the P450c17 inhibitor, ketoconazole (10 nmol) significantly inhibited the CCI-induced increase in GFAP-immunoreactivity, but had no effect on the CCI-induced increase in Iba-1-immunoreactivity. In addition, intrathecal administration of ketoconazole significantly inhibited the CCI-induced increase in p38 phosphorylation, while the levels of ERK and JNK phosphorylation remained unchanged. The CCI-induced development of mechanical allodynia was attenuated by administration of either ketoconazole (10 nmol) or the p38 MAPK inhibitor, SB203580 (5 nmol). Administration of a sub-effective dose of SB203580 (0.5 nmol) potentiated the pharmacological effect of ketoconazole (1 nmol) on spinal GFAP-immunostaining, as well as, the development of mechanical allodynia following CCI. Collectively these data suggest that spinal P450c17 activates astrocytes via p38 phosphorylation, ultimately leading to the development of mechanical allodynia in a model of peripheral neuropathy.


Assuntos
Astrócitos/enzimologia , Neuralgia/enzimologia , Neuralgia/patologia , Medula Espinal/patologia , Esteroide 17-alfa-Hidroxilase/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/patologia , Constrição Patológica , Modelos Animais de Doenças , Hiperalgesia/complicações , Hiperalgesia/patologia , Imidazóis/farmacologia , Cetoconazol/administração & dosagem , Cetoconazol/farmacologia , Vértebras Lombares/enzimologia , Vértebras Lombares/patologia , Masculino , Camundongos , Microglia/efeitos dos fármacos , Microglia/patologia , Fosforilação/efeitos dos fármacos , Piridinas/farmacologia , Corno Dorsal da Medula Espinal/enzimologia , Corno Dorsal da Medula Espinal/patologia , Esteroide 17-alfa-Hidroxilase/metabolismo
3.
Anesth Analg ; 129(2): 578-586, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-29933273

RESUMO

BACKGROUND: Although endogenous analgesia plays an important role in controlling pain states, chronic pain patients exhibit decreased endogenous analgesia compared to healthy individuals. In rats, noxious stimulus-induced analgesia (NSIA), which is an indicator of endogenous analgesia, diminished 6 weeks after spinal nerve ligation (SNL6W). A recent study in rats with deleted noradrenergic fibers demonstrated that the noradrenergic fibers were essential to NSIA. It has also been reported that brain-derived neurotrophic factor increased spinal noradrenergic fibers. Therefore, this study examined the effect of TrkB activation, which is the receptor for brain-derived neurotrophic factor, on impaired NSIA in SNL6W rats. In addition, we also examined the effect of endogenous analgesia on acute incisional pain. METHODS: After 5 daily intraperitoneal injections of 7,8-dihydroxyflavone (7,8-DHF, TrkB agonist, 5 mg/kg), NSIA was examined by measuring the withdrawal threshold increment in the left (contralateral to nerve ligation) hindpaw at 30 minutes after capsaicin injection (250 µg) in the forepaw. K252a (TrkB antagonist, 2 µg) was administrated intrathecally for 5 days. Idazoxan (α2 adrenoceptor antagonist, 30 µg), atropine (muscarinic antagonist, 30 µg), and propranolol (nonselective ß adrenoceptor antagonist, 30 µg) were administered intrathecally for 15 minutes before capsaicin injection. Microdialysis and immunohistochemistry were performed to examine the noradrenergic plasticity in the spinal dorsal horn. A hindpaw incision was performed on the left (contralateral to nerve ligation) hindpaw. Data were analyzed by 1-way analyses of variance or 2-way repeated-measures 1-way analysis of variance followed by a Student t test with Bonferroni correction. RESULTS: Five daily intraperitoneal injections of 7,8-DHF restored the attenuated NSIA in SNL6W rats (n = 7, P = .002; estimated treatment effect [95% CI]: 62.9 [27.0-98.7] g), with this effect blocked by 5 daily intrathecal coadministrations of K252a (n = 6, P < .001; -57.8 [-78.3 to -37.2] g). This effect was also inhibited by a single intrathecal administration of idazoxan (n = 8, P < .001; -61.6 [-92.4 to -30.9] g) and atropine (n = 8, P = .003; -52.6 [-73.3 to -31.9] g), but not by propranolol. Furthermore, 7,8-DHF increased the noradrenergic fiber in the spinal dorsal horn and the noradrenaline release in response to the capsaicin injection in the forepaw in SNL6W rats. In addition, repeated injections of 7,8-DHF prevented delayed recovery from incisional pain in SNL6W rats. CONCLUSIONS: Spinal activation of TrkB may recover the attenuated endogenous analgesia by improving the adrenergic plasticity, thereby leading to prevention of pain prolongation after surgery.


Assuntos
Analgésicos/farmacologia , Flavonas/farmacologia , Neuralgia/tratamento farmacológico , Limiar da Dor/efeitos dos fármacos , Receptor trkB/agonistas , Corno Dorsal da Medula Espinal/efeitos dos fármacos , Fibras Adrenérgicas/efeitos dos fármacos , Fibras Adrenérgicas/enzimologia , Animais , Modelos Animais de Doenças , Ativação Enzimática , Masculino , Neuralgia/enzimologia , Neuralgia/fisiopatologia , Plasticidade Neuronal/efeitos dos fármacos , Norepinefrina/metabolismo , Ratos Sprague-Dawley , Receptor trkB/metabolismo , Transdução de Sinais , Corno Dorsal da Medula Espinal/enzimologia , Corno Dorsal da Medula Espinal/fisiopatologia
4.
Brain Res ; 1646: 570-579, 2016 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-27378584

RESUMO

Lamina X of the spinal cord is a functionally diverse area with roles in locomotion, autonomic control and processing of mechano and nociceptive information. It is also a neurochemically diverse region. However, the different populations of cells in lamina X remain to be fully characterised. To determine the co-localisation of the enzymes responsible for the production of GABA and acetylcholine (which play major roles in the spinal cord) in lamina X of the adult and juvenile mouse, we used a transgenic mouse expressing green fluorescent protein (GFP) in glutamate decarboxylase 67 (GAD67) neurons, combined with choline acetyltransferase (ChAT) immunohistochemistry. ChAT-immunoreactive (IR) and GAD67-GFP containing neurons were observed in lamina X of both adult and juvenile mice and in both age groups a population of cells containing both ChAT-IR and GAD67-GFP were observed in lumbar, thoracic and cervical spinal cord. Such dual labelled cells were predominantly located ventral to the central canal. Immunohistochemistry for vesicular acetylcholine transporter (VAChT) and GAD67 revealed a small number of double labelled terminals located lateral, dorsolateral and ventrolateral to the central canal. This study therefore describes in detail a population of ChAT-IR/GAD67-GFP neurons predominantly ventral to the central canal of the cervical, thoracic and lumbar spinal cord of adult and juvenile mice. These cells potentially correspond to a sub-population of the cholinergic central canal cluster cells which may play a unique role in controlling spinal cord circuitry.


Assuntos
Colina O-Acetiltransferase/metabolismo , Glutamato Descarboxilase/metabolismo , Neurônios/enzimologia , Corno Dorsal da Medula Espinal/enzimologia , Acetilcolina/metabolismo , Animais , Feminino , Proteínas de Fluorescência Verde/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Ácido gama-Aminobutírico/metabolismo
5.
Mol Pain ; 122016.
Artigo em Inglês | MEDLINE | ID: mdl-27030712

RESUMO

BACKGROUND: Physical exercise effectively attenuates neuropathic pain, and multiple events including the inhibition of activated glial cells in the spinal dorsal horn, activation of the descending pain inhibitory system, and reductions in pro-inflammatory cytokines in injured peripheral nerves may contribute to exercise-induced hypoalgesia. Since fewer GABAergic hypoalgesic interneurons exist in the dorsal horn in neuropathic pain model animals, the recovery of impaired GABAergic inhibition in the dorsal horn may improve pain behavior. We herein determined whether the production of gamma-aminobutyric acid (GABA) and glutamic acid decarboxylase (GAD) in the dorsal horn is restored by treadmill running and contributes to exercise-induced hypoalgesia in neuropathic pain model mice. C57BL/6 J mice underwent partial sciatic nerve ligation (PSL). PSL-Runner mice ran on a treadmill at 7 m/min for 60 min/day, 5 days/week, from two days after PSL. RESULTS: Mechanical allodynia and heat hyperalgesia developed in PSL-Sedentary mice but were significantly attenuated in PSL-Runner mice. PSL markedly decreased GABA and GAD65/67 levels in neuropils in the ipsilateral dorsal horn, while treadmill running inhibited these reductions. GABA+ neuronal nuclei+ interneuron numbers in the ipsilateral dorsal horn were significantly decreased in PSL-Sedentary mice but not in PSL-Runner mice. Pain behavior thresholds positively correlated with GABA and GAD65/67 levels and GABAergic interneuron numbers in the ipsilateral dorsal horns of PSL-Sedentary and -Runner mice. CONCLUSIONS: Treadmill running prevented PSL-induced reductions in GAD65/67 production, and, thus, GABA levels may be retained in interneurons and neuropils in the superficial dorsal horn. Therefore, improvements in impaired GABAergic inhibition may be involved in exercise-induced hypoalgesia.


Assuntos
Glutamato Descarboxilase/metabolismo , Neuralgia/enzimologia , Neuralgia/patologia , Condicionamento Físico Animal , Corno Dorsal da Medula Espinal/enzimologia , Ácido gama-Aminobutírico/metabolismo , Animais , Comportamento Animal , Modelos Animais de Doenças , Masculino , Camundongos Endogâmicos C57BL , Neuralgia/fisiopatologia , Limiar da Dor , Corno Dorsal da Medula Espinal/patologia
6.
Mol Pain ; 11: 49, 2015 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-26272725

RESUMO

BACKGROUND: The rostral anterior cingulate cortex (rACC) has been implicated in the negative affective response to injury, and importantly, it has been shown that activation of extracellular signal-regulated kinase (ERK) signaling in the rACC contributes to the full expression of the affective component of pain in rodents. In this study, we investigated whether administration of anesthesia at the time of injury could reduce phosphorylated-ERK (PERK) expression in the rACC, which might eliminate the negative affective component of noxious stimulation. Intraplantar hindpaw formalin stimulation, an aversive event in the awake animal, was given with or without general isoflurane anesthesia, and PERK expression was subsequently quantified in the rACC using immunohistochemistry. Furthermore, as numerous studies have demonstrated the importance of spinal ERK signaling in the regulation of nociceptive behaviour, we also examined PERK in the superficial dorsal horn of the spinal cord. FINDINGS: Formalin injection with and without short-term (<10 min) general isoflurane anesthesia induced the same level of PERK expression in spinal cord laminae I-II. However, PERK expression was significantly inhibited across all laminae of the rACC in animals anesthetized during formalin injection. The effect of anesthesia was such that levels of PERK were the same in formalin and sham treated anesthesized animals. CONCLUSIONS: This study is the first to demonstrate that isoflurane anesthesia can inhibit formalin-induced PERK in the rACC and therefore might eliminate the unpleasantness of restraint associated with awake hindpaw injection.


Assuntos
Anestesia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Formaldeído/farmacologia , Giro do Cíngulo/enzimologia , Medula Espinal/enzimologia , Animais , Ativação Enzimática/efeitos dos fármacos , Giro do Cíngulo/efeitos dos fármacos , Masculino , Fosforilação/efeitos dos fármacos , Ratos Sprague-Dawley , Medula Espinal/efeitos dos fármacos , Corno Dorsal da Medula Espinal/efeitos dos fármacos , Corno Dorsal da Medula Espinal/enzimologia , Fatores de Tempo
7.
Mol Pain ; 11: 46, 2015 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-26255228

RESUMO

BACKGROUND: Koumine is an alkaloid monomer found abundantly in Gelsemium plants. It has been shown to reverse thermal hyperalgesia and mechanical allodynia induced by sciatic nerve chronic constriction injury (CCI) in rats in a dose-dependent manner. Interestingly, this effect is mediated by elevated allopregnanolone levels in the spinal cord (SC). Since 3α-hydroxysteroid oxidoreductase (3α-HSOR), the key synthetase of allopregnanolone, is responsible for allopregnanolone upregulation in the SC, the objective of the present study was to investigate the role of its expression in the SC in koumine-induced analgesia using a rat model of neuropathic pain following peripheral nerve injury. RESULTS: Time-course investigations of immunohistochemistry and real-time polymerase chain reaction revealed that the immunoreactivity and mRNA expression of 3α-HSOR markedly increased in a time-dependent manner in the SC of koumine-treated CCI rats. Furthermore, 3α-HSOR activity in the SC of koumine-treated CCI rats increased by 15.8% compared to the activity in untreated CCI rats. Intrathecal injection of medroxyprogesterone acetate, a selective 3α-HSOR inhibitor, reversed the analgesic effect of koumine on CCI-induced mechanical pain perception. Our results confirm that koumine alleviates neuropathic pain in rats with CCI by enhancing 3α-HSOR mRNA expression and bioactivity in the SC. CONCLUSION: This study demonstrates that 3α-HSOR is an important molecular target of koumine for alleviating neuropathic pain. Koumine may prove a promising compound for the development of novel analgesic agents effective against intractable neuropathic pain.


Assuntos
Alcaloides Indólicos/uso terapêutico , Neuralgia/tratamento farmacológico , Neuralgia/enzimologia , Medula Espinal/enzimologia , 3-alfa-Hidroxiesteroide Desidrogenase (B-Específica)/antagonistas & inibidores , 3-alfa-Hidroxiesteroide Desidrogenase (B-Específica)/genética , 3-alfa-Hidroxiesteroide Desidrogenase (B-Específica)/metabolismo , Analgésicos/farmacologia , Analgésicos/uso terapêutico , Animais , Biocatálise/efeitos dos fármacos , Doença Crônica , Constrição , Modelos Animais de Doenças , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Hiperalgesia/complicações , Hiperalgesia/tratamento farmacológico , Hiperalgesia/patologia , Imuno-Histoquímica , Alcaloides Indólicos/administração & dosagem , Alcaloides Indólicos/química , Alcaloides Indólicos/farmacologia , Injeções Subcutâneas , Masculino , Acetato de Medroxiprogesterona/farmacologia , Neuralgia/complicações , Neuralgia/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/enzimologia , Nervo Isquiático/patologia , Medula Espinal/efeitos dos fármacos , Medula Espinal/patologia , Corno Dorsal da Medula Espinal/efeitos dos fármacos , Corno Dorsal da Medula Espinal/enzimologia , Corno Dorsal da Medula Espinal/patologia
8.
Morfologiia ; 147(2): 32-7, 2015.
Artigo em Russo | MEDLINE | ID: mdl-26234037

RESUMO

The aim of this study was to examine the distribution of cholinergic and nitroxidergic neurons in the spinal cord (SC) of adult and newborn rats. Using immunohistochemical demonstration of choline acetyltransferase (ChAT) and nitric oxide synthase (NOS), cervical portions of SC were studied in newborn (n=5) and adult (n=5) Wistar rats. It was found that ChAT-positive neurons were localized in the anterior horns of the SC, while individual cells were located in of SC posterior horns, in the central gray matter and at the boundary of VI-VII Rexed laminae. Nitroxidergic neurons were located in the superficial layers of SC posterior horns of grey matter, in the central gray matter and in the area of VI-VII Rexed laminae. It is found that SC of newborn and adult rats contained cholinergic neurons expressing NOS. Detection of cells containing both enzymes already at postnatal Day 1, suggests that they were formed in rat SC during prenatal ontogenesis


Assuntos
Neurônios Colinérgicos , Neurônios Nitrérgicos , Óxido Nítrico Sintase Tipo I/metabolismo , Corno Dorsal da Medula Espinal , Corno Lateral da Medula Espinal , Animais , Animais Recém-Nascidos , Neurônios Colinérgicos/citologia , Neurônios Colinérgicos/enzimologia , Neurônios Nitrérgicos/citologia , Neurônios Nitrérgicos/enzimologia , Ratos , Ratos Wistar , Corno Dorsal da Medula Espinal/citologia , Corno Dorsal da Medula Espinal/enzimologia , Corno Lateral da Medula Espinal/citologia , Corno Lateral da Medula Espinal/enzimologia
9.
Mol Pain ; 11: 36, 2015 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-26104415

RESUMO

BACKGROUND: The descending antinociceptive system (DAS) is thought to play crucial roles in the antinociceptive effect of spinal cord stimulation (SCS), especially through its serotonergic pathway. The nucleus raphe magnus (NRM) in the rostral ventromedial medulla is a major source of serotonin [5-hydroxytryptamine (5-HT)] to the DAS, but the role of the dorsal raphe nucleus (DRN) in the ventral periaqueductal gray matter is still unclear. Moreover, the influence of the noradrenergic pathway is largely unknown. In this study, we evaluated the involvement of these serotonergic and noradrenergic pathways in SCS-induced antinociception by behavioral analysis of spinal nerve-ligated (SNL) rats. We also investigated immunohistochemical changes in the DRN and locus coeruleus (LC), regarded as the adrenergic center of the DAS, and expression changes of synthetic enzymes of 5-HT [tryptophan hydroxylase (TPH)] and norepinephrine [dopamine ß-hydroxylase (DßH)] in the spinal dorsal horn. RESULTS: Intrathecally administered methysergide, a 5-HT1- and 5-HT2-receptor antagonist, and idazoxan, an α2-adrenergic receptor antagonist, equally abolished the antinociceptive effect of SCS. The numbers of TPH-positive serotonergic and phosphorylated cyclic AMP response element binding protein (pCREB)-positive neurons and percentage of pCREB-positive serotonergic neurons in the DRN significantly increased after 3-h SCS. Further, the ipsilateral-to-contralateral immunoreactivity ratio of DßH increased in the LC of SNL rats and reached the level seen in naïve rats, even though the number of pCREB-positive neurons in the LC was unchanged by SNL and SCS. Moreover, 3-h SCS did not increase the expression levels of TPH and DßH in the spinal dorsal horn. CONCLUSIONS: The serotonergic and noradrenergic pathways of the DAS are involved in the antinociceptive effect of SCS, but activation of the DRN might primarily be responsible for this effect, and the LC may have a smaller contribution. SCS does not potentiate the synthetic enzymes of 5HT and norepinephrine in the neuropathic spinal cord.


Assuntos
Nociceptividade , Estimulação da Medula Espinal/métodos , Nervos Espinhais/lesões , Neurônios Adrenérgicos/efeitos dos fármacos , Neurônios Adrenérgicos/metabolismo , Analgésicos/farmacologia , Animais , Western Blotting , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Dopamina beta-Hidroxilase/metabolismo , Núcleo Dorsal da Rafe/efeitos dos fármacos , Núcleo Dorsal da Rafe/metabolismo , Idazoxano/farmacologia , Locus Cerúleo/efeitos dos fármacos , Locus Cerúleo/metabolismo , Metisergida/farmacologia , Nociceptividade/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Ratos , Neurônios Serotoninérgicos/efeitos dos fármacos , Neurônios Serotoninérgicos/metabolismo , Corno Dorsal da Medula Espinal/efeitos dos fármacos , Corno Dorsal da Medula Espinal/enzimologia , Nervos Espinhais/efeitos dos fármacos , Nervos Espinhais/patologia , Triptofano Hidroxilase/metabolismo
10.
Mol Pain ; 11: 35, 2015 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-26093674

RESUMO

BACKGROUND: Neuropathic pain and sensory abnormalities are a debilitating secondary consequence of spinal cord injury (SCI). Maladaptive structural plasticity is gaining recognition for its role in contributing to the development of post SCI pain syndromes. We previously demonstrated that excitotoxic induced SCI dysesthesias are associated with enhanced dorsal root ganglia (DRG) neuronal outgrowth. Although glycogen synthase kinase-3ß (GSK-3ß) is a known intracellular regulator neuronal growth, the potential contribution to primary afferent growth responses following SCI are undefined. We hypothesized that SCI triggers inhibition of GSK-3ß signaling resulting in enhanced DRG growth responses, and that PI3K mediated activation of GSK-3ß can prevent this growth and the development of at-level pain syndromes. RESULTS: Excitotoxic SCI using intraspinal quisqualic acid (QUIS) resulted in inhibition of GSK-3ß in the superficial spinal cord dorsal horn and adjacent DRG. Double immunofluorescent staining showed that GSK-3ß(P) was expressed in DRG neurons, especially small nociceptive, CGRP and IB4-positive neurons. Intrathecal administration of a potent PI3-kinase inhibitor (LY294002), a known GSK-3ß activator, significantly decreased GSK-3ß(P) expression levels in the dorsal horn. QUIS injection resulted in early (3 days) and sustained (14 days) DRG neurite outgrowth of small and subsequently large fibers that was reduced with short term (3 days) administration of LY294002. Furthermore, LY294002 treatment initiated on the date of injury, prevented the development of overgrooming, a spontaneous at-level pain related dysesthesia. CONCLUSIONS: QUIS induced SCI resulted in inhibition of GSK-3ß in primary afferents and enhanced at-level DRG intrinsic growth (neurite elongation and initiation). Early PI3K mediated activation of GSK-3ß attenuated QUIS-induced DRG neurite outgrowth and prevented the development of at-level dysesthesias.


Assuntos
Quinase 3 da Glicogênio Sintase/metabolismo , Neurônios Aferentes/enzimologia , Neurônios Aferentes/patologia , Neurotoxinas/toxicidade , Parestesia/complicações , Fosfatidilinositol 3-Quinases/metabolismo , Traumatismos da Medula Espinal/complicações , Animais , Cromonas/administração & dosagem , Cromonas/farmacologia , Ativação Enzimática/efeitos dos fármacos , Gânglios Espinais/efeitos dos fármacos , Gânglios Espinais/enzimologia , Gânglios Espinais/patologia , Glicogênio Sintase Quinase 3 beta , Injeções Espinhais , Masculino , Morfolinas/administração & dosagem , Morfolinas/farmacologia , Neuritos/efeitos dos fármacos , Neuritos/patologia , Neurônios Aferentes/efeitos dos fármacos , Nociceptividade/efeitos dos fármacos , Parestesia/enzimologia , Parestesia/patologia , Inibidores de Proteínas Quinases/farmacologia , Ácido Quisquálico , Ratos Long-Evans , Corno Dorsal da Medula Espinal/efeitos dos fármacos , Corno Dorsal da Medula Espinal/enzimologia , Corno Dorsal da Medula Espinal/patologia , Traumatismos da Medula Espinal/enzimologia , Traumatismos da Medula Espinal/patologia
11.
Neuroscience ; 291: 272-8, 2015 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-25701711

RESUMO

Whether visceral organ cross-sensitization is involved in endometriosis-associated pain remains elusive. Previous studies have shown that visceral noxious stimuli may trigger a cascade of signal transductions in the rostral ventromedial medulla (RVM) via the spinal dorsal column (DC) pathway and the RVM plays a critical role in the descending control of visceral nociception. In the current study, we hypothesized that the p38 mitogen-activated protein kinase (MAPK) activation in the RVM by noxious visceral inputs from ectopic growths via the DC was involved in the development of pelvic organ cross-sensitization in established endometriosis. A rat model of experimental endometriosis was established. To examine ectopic growths-to-colon cross-sensitization, graded colorectal distention (CRD) was performed and abdominal withdrawal reflex (AWR) scores were recorded in female rats at 8weeks after the uterine or fat (control) auto-transplantation. Western blot study was carried out to examine the phosphorylated form and the total level of p38 MAPK protein in the RVM. Our results showed that lesions of bilateral DCs immediately following uterine or fat auto-transplantation in female rats significantly attenuated the later development of ectopic growths-to-colon cross-sensitization and the increased p38 MAPK activation in the RVM, as compared to sham DC lesions. Furthermore, intra-RVM microinjection of a p38 MAPK inhibitor (SB 203580), but not vehicle, in female rats with established endometriosis significantly attenuated ectopic growths-to-colon cross-sensitization and the increased activation of p38 MAPK in the RVM. These findings suggest that the noxious inputs from ectopic growths may activate p38 MAPK in the RVM via the DC, which may contribute to the development of ectopic growths-to-colon cross-sensitization in established endometriosis.


Assuntos
Colo/enzimologia , Endometriose/enzimologia , Bulbo/enzimologia , Dor Visceral/enzimologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Tecido Adiposo/transplante , Animais , Modelos Animais de Doenças , Feminino , Imidazóis/farmacologia , Bulbo/efeitos dos fármacos , Limiar da Dor/efeitos dos fármacos , Limiar da Dor/fisiologia , Fosforilação/efeitos dos fármacos , Estimulação Física , Pressão , Inibidores de Proteínas Quinases/farmacologia , Piridinas/farmacologia , Ratos Sprague-Dawley , Corno Dorsal da Medula Espinal/enzimologia , Transplante Autólogo , Útero/transplante , Dor Visceral/tratamento farmacológico , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores
12.
Tissue Cell ; 47(1): 55-60, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25497384

RESUMO

The aim of the present study was to investigate whether the phosphorylation of ERK1/2 in the rat lumbar dorsal horn and in the parvocellularis part of the paraventricular nucleus can be used to visualize neuronal activity. pERK1/2 fluorescence-immunohistochemistry is specifically suited to mirror neuronal activity in the pain pathway following an acute noxious stimulation. The rat hind paw was either stimulated by noxious heat or by a sequence of mustard oil and noxious heat. Two and 10 min after the thermal stimulation a 3-4-fold increase in cells with pERK1/2 immunoreactivity was observed in lamina I/II of the L3-L5 dorsal horn. The combination of mustard oil with heat led to a 5-6-fold increase in the pERK1/2 signal. The pERK1/2 immunoreactivity in the parvocellularis part of the paraventricular nucleus increased by 2-fold following the heat stimulus, with no further increase following the sequential mustard oil and heat stimulus. A pretreatment with the opioid analgesic morphine or the NMDA antagonist MK-801 markedly attenuated ERK1/2 phosphorylation in both areas of the pain pathway. The present findings support the concept that the pERK1/2 immunofluorescence signal can be used as a quantitative marker for sensitization or inhibition in the pain pathway at spinal and hypothalamic level.


Assuntos
Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Neurônios/enzimologia , Dor/genética , Núcleo Hipotalâmico Paraventricular/enzimologia , Corno Dorsal da Medula Espinal/enzimologia , Animais , Imunofluorescência , Temperatura Alta , Masculino , Morfina/administração & dosagem , Mostardeira/toxicidade , Neurônios/patologia , Dor/induzido quimicamente , Dor/patologia , Núcleo Hipotalâmico Paraventricular/patologia , Fosforilação , Óleos de Plantas/toxicidade , Ratos , Ratos Sprague-Dawley , Corno Dorsal da Medula Espinal/patologia
13.
PLoS One ; 9(3): e89149, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24598860

RESUMO

This study was designed to investigate the anti-inflammatory and anti-nociceptive activity of the methanol extract from the aerial part of Phlomis younghusbandii (MEAP) and to explore the possible related mechanisms. Anti-inflammatory effects of MEAP were evaluated by using the ear edema test induced by dimethylbenzene and vascular permeability test induced by acetic acid. Anti-nociceptive activities of MEAP were evaluated by the chemical nociception in models of acetic acid-induced writhing and formalin-induced hind paw licking, and by the thermal nociception in hot plate tests. Mechanisms of MEAP activities also were explored by evaluating expression levels of TNF-α, IL-6 and iNOS induced by LPS using real-time fluorogenic PCR and expression of COX-2 using Western blotting and an open-field test. The results indicated that the MEAP administered orally could significantly decrease ear edema induced by dimethylbenzene and increase vascular permeability induced by acetic acid. Additionally, the nociceptions induced by acetic acid and formalin were significantly inhibited. The anti-nociceptive effect could not be decreased by naloxone in the formalin test, and MEAP did not affect the normal autonomic activities of mice. Expression levels of pro-inflammatory cytokines (TNF-α, IL-6, iNOS) induced by LPS were decreased obviously by treatment with MEAP. Furthermore, COX-2 expression in the spinal dorsal horns of the pain model mice induced by formalin was significantly down-regulated by MEAP. In conclusion, MEAP has significant anti-inflammatory and antinociceptive activities, and the mechanisms may be related to the down-regulated expression of TNF-α, IL-6, iNOS and COX-2.


Assuntos
Analgésicos/farmacologia , Anti-Inflamatórios/farmacologia , Phlomis/química , Componentes Aéreos da Planta/química , Extratos Vegetais/farmacologia , Analgésicos/isolamento & purificação , Animais , Anti-Inflamatórios/isolamento & purificação , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Ciclo-Oxigenase 2/metabolismo , Regulação para Baixo/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Indometacina/farmacologia , Interleucina-6/metabolismo , Lipopolissacarídeos/farmacologia , Metanol/química , Camundongos Endogâmicos ICR , Morfina/farmacologia , Nociceptividade/efeitos dos fármacos , Extratos Vegetais/isolamento & purificação , Solventes/química , Corno Dorsal da Medula Espinal/efeitos dos fármacos , Corno Dorsal da Medula Espinal/enzimologia , Fator de Necrose Tumoral alfa/metabolismo
14.
Neuropathol Appl Neurobiol ; 40(4): 452-63, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23899306

RESUMO

AIMS: Recent studies suggested that the development of neuropathic pain associated with neural injury may be partly due to up-regulation of cyclooxygenase (COX) in the central nervous system. However, the cellular sources of COX-1 and COX-2 up-regulation following nerve injury are unclear. METHODS: We investigated the spinal cellular sources of COX-1 and COX-2 in association with allodynia following L5 spinal nerve ligation (SNL). RESULTS: Post-SNL pain-related behaviour was shown by increased sensitivity to mechanical stimulation. There was a significant increase in both COX-1 and COX-2 immunoreactivity (P < 0.01) on the ipsilateral side of spinal dorsal horn. Double immunofluorescence labelling demonstrated that COX-1 immunoreactive cells colocalized chiefly with dorsal horn neuronal nuclei and microglia, whereas COX-2 was expressed in neuronal cytoplasm. CONCLUSION: These findings demonstrate that while spinal dorsal horn neurones are important source of COX-1 and COX-2 after nerve injury, microglia also contribute to the pathogenesis of neuropathic pain, partly by producing additional COX-1.


Assuntos
Ciclo-Oxigenase 1/metabolismo , Ciclo-Oxigenase 2/metabolismo , Neuralgia/enzimologia , Neuroglia/enzimologia , Neurônios/enzimologia , Corno Dorsal da Medula Espinal/enzimologia , Animais , Hiperalgesia/enzimologia , Ligadura , Masculino , Ratos , Ratos Sprague-Dawley , Nervos Espinhais/lesões , Regulação para Cima
15.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 38(7): 686-90, 2013 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-23908078

RESUMO

OBJECTIVE: To investigate the changes of cyclooxygenase-2 (COX-2) expression in the spinal cord dorsal horn after intrathecal a specific p38MAPK inhibitor-SB203580 on neuropathic pain in rats induced by chronic constrictive injury (CCI) to the sciatic nerve. METHODS: Twenty-four male SD rats after intrathecal catheter placement were randomly divided into 4 groups: a sham group with sham surgery, the neuropathic pain model of a NS group, a DMSO group and an SB group were established by CCI to sciatic nerve. NS or DMSO or SB203580 was injected IT NS or 2%DMSO or SB203580 twice a day for 5 consecutive days starting at 6th day when the model of chronic constrictive injury was established. Mechanical stimuli were measured before the surgery and on 1st, 3rd, 5th, 7th, 9th, and 11th day after the surgery. Then all rats were sacrificed and the lumbar segment of spinal cord was removed to determine the COX-2 expression in the dorsal horn by immunocytochemistry. RESULTS: Day 1 to 11 after the surgery, the threshold to mechanical on the surgery side was significantly lower in the NS group and the DMSO group than in the sham group. Day 7 to 11 after the sugery, the threshold to mechanical on the surgery side was significantly lower in the SB group than in the NS group and the DMSO group. The expression of spinal COX-2 was higher in the NS group and the DMSO group than in the sham group, but lower in the SB group than in NS group and the DMSO group. CONCLUSION: Intrathecal administration of SB203580 has significant analgesic effect in the CCI rat model. Expression of COX-2 is significantly reduced when p38MAPK is inhibited by intrathecal SB203580, and p38MAPK stimulation is essential for COX-2 expression.


Assuntos
Ciclo-Oxigenase 2/metabolismo , Imidazóis/administração & dosagem , Neuralgia/enzimologia , Piridinas/administração & dosagem , Corno Dorsal da Medula Espinal/enzimologia , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Animais , Ciclo-Oxigenase 2/genética , Imidazóis/farmacologia , Injeções Espinhais , Masculino , Neuralgia/fisiopatologia , Piridinas/farmacologia , Ratos , Ratos Sprague-Dawley , Medula Espinal/enzimologia
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