RESUMO
BACKGROUND: Parkinson's disease (PD) is characterized by Lewy body and neurite pathology associated with dopamine terminal dysfunction. Clinically, it is associated with motor slowing, rigidity, and tremor. Postural instability and pain are also features. Physical exercise benefits PD patients - possibly by promoting neuroplasticity including synaptic regeneration. OBJECTIVES: In a parkinsonian rat model, we test the hypotheses that exercise: (a) increases synaptic density and reduces neuroinflammation and (b) lowers the nociceptive threshold by increasing µ-opioid receptor expression. METHODS: Brain autoradiography was performed on rats unilaterally injected with either 6-hydroxydopamine (6-OHDA) or saline and subjected to treadmill exercise over 5 weeks. [3H]UCB-J was used to measure synaptic vesicle glycoprotein 2A (SV2A) density. Dopamine D2/3 receptor and µ-opioid receptor availability were assessed with [3H]Raclopride and [3H]DAMGO, respectively, while neuroinflammation was detected with the 18kDA translocator protein (TSPO) marker [3H]PK11195. The nociceptive threshold was determined prior to and throughout the exercise protocol. RESULTS: We confirmed a dopaminegic deficit with increased striatal [3H]Raclopride D2/3 receptor availability and reduced nigral tyrosine hydroxylase immunoreactivity in the ipsilateral hemisphere of all 6-OHDA-injected rats. Sedentary rats lesioned with 6-OHDA showed significant reduction of ipsilateral striatal and substantia nigra [3H]UCB-J binding while [3H]PK11195 showed increased ipsilateral striatal neuroinflammation. Lesioned rats who exercised had higher levels of ipsilateral striatal [3H]UCB-J binding and lower levels of neuroinflammation compared to sedentary lesioned rats. Striatal 6-OHDA injections reduced thalamic µ-opioid receptor availability but subsequent exercise restored binding. Exercise also raised thalamic and hippocampal SV2A synaptic density in 6-OHDA lesioned rats, accompanied by a rise in nociceptive threshold. CONCLUSION: These data suggest that treadmill exercise protects nigral and striatal synaptic integrity in a rat lesion model of PD - possibly by promoting compensatory mechanisms. Exercise was also associated with reduced neuroinflammation post lesioning and altered opioid transmission resulting in an increased nociceptive threshold.
Assuntos
Encéfalo/metabolismo , Transtornos Parkinsonianos/metabolismo , Transtornos Parkinsonianos/terapia , Condicionamento Físico Animal/fisiologia , Sinapses/metabolismo , Animais , Encéfalo/efeitos dos fármacos , Teste de Esforço/métodos , Masculino , Oxidopamina/toxicidade , Transtornos Parkinsonianos/induzido quimicamente , Condicionamento Físico Animal/métodos , Ratos , Ratos Wistar , Sinapses/efeitos dos fármacosRESUMO
Various animal models have been employed to understand the pathogenic mechanism of neuropathic pain. Nitric oxide (NO) is an important molecule in nociceptive transmission and is involved in neuropathic pain. However, its mechanistic actions remain unclear. The aim of this study was to better understand the involvement of neuronal and inducible isoforms of nitric oxide synthase (nNOS and iNOS) in neuropathic pain induced by chronic constriction injury (CCI) of the sciatic nerve in rats. We evaluated pain sensitivity (mechanical withdrawal thresholds using Randall and Selitto, and von Frey tests, and thermal withdrawal thresholds using Hargreaves test) prior to CCI surgery, 14 days post CCI and after intrathecal injections of selective nNOS or iNOS inhibitors. We also evaluated the distribution of NOS isozymes in the spinal cord and dorsal root ganglia (DRG) by immunohistochemistry, synthesis of iNOS and nNOS by Western blot, and NO production using fluorescent probe DAF-2 DA (DA). Our results showed higher number of nNOS and iNOS-positive neurons in the spinal cord and DRG of CCI compared to sham rats, and their reduction in CCI rats after treatment with selective inhibitors compared to non-treated groups. Western blot results also indicated reduced expression of nNOS and iNOS after treatment with selective inhibitors. Furthermore, both inhibitors reduced CCI-evoked mechanical and thermal withdrawal thresholds but only nNOS inhibitor was able to efficiently lower mechanical withdrawal thresholds using von Frey test. In addition, we observed higher NO production in the spinal cord and DRG of injured rats compared to control group. Our study innovatively shows that nNOS may strongly modulate nociceptive transmission in rats with neuropathic pain, while iNOS may partially participate in the development of nociceptive responses. Thus, drugs targeting nNOS for neuropathic pain may represent a potential therapeutic strategy.
Assuntos
Gânglios Espinais/metabolismo , Neuralgia/metabolismo , Óxido Nítrico/metabolismo , Nervo Isquiático/metabolismo , Animais , Hiperalgesia/tratamento farmacológico , Masculino , Óxido Nítrico Sintase Tipo II/metabolismo , Ratos Wistar , Medula Espinal/metabolismoRESUMO
Chronic constriction injury (CCI) of infraorbital nerve (IoN) results in whisker pad mechanical allodynia in rats and activation glial cells contributing to the development of orofacial pain. Whisker pad mechanical allodynia (von Frey stimuli) was tested pre and postoperatively and conducted during the treatment time. Photobiomodulation (PBM) and vitamins B complex (VBC) has been demonstrated therapeutic efficacy in ameliorate neuropathic pain. The aim of this study was to evaluate the antinociceptive effect of PBM, VBC or the combined treatment VBC â+ âPBM on orofacial pain due to CCI-IoN. Behavioral and molecular approaches were used to analyses nociception, cellular and neurochemical alterations. CCI-IoN caused mechanical allodynia and cellular alterations including increased expression of glial fibrillary acid protein (GFAP) and ionized calcium binding adaptor molecule 1 (Iba-1), administration of VBC (B1/B6/B12 at 180/180/1.8 âmg/kg, s.c., 5 times all long 10 sessions) and PBM therapy (904 ânm, power of 75Wpico, average power of 0.0434 âW, pulse frequency of 9500 âHz, area of the beam 0.13 âcm2, 18 âs duration, energy density 6 âJ/cm2, with an energy per point of 0.78 âJ for 10 sessions) or their combination presented improvement of the nociceptive behavior and decreased expression of GFAP and Iba-1. Additionally, CCI-IoN rats exhibited an upregulation of IL1ß, IL6 and TNF-α expression and all treatments prevented this upregulation and also increased IL10 expression. Overall, the present results highlight the pain reliever effect of VBC or PBM alone or in combination, through the modulation of glial cells and cytokines expression in the spinal trigeminal nucleus of rats.
RESUMO
In addition to motor symptoms, Parkinson's disease (PD) presents high prevalence of painful symptoms responsible for worsening quality of life of PD patients. Physical exercise can improve such painful symptoms. This study evaluated the effects of exercise on nociceptive threshold using an unilateral rat model of PD, as well as the role played by cannabinoid and opioid receptors in areas responsible for pain pathways. For PD induction, Wistar rats were injected with 6-OHDA. 15â¯days after, rats either remained sedentary or were forced to exercise three times a week for 40â¯min. Motor and nociceptive behaviors were evaluated through cylinder and mechanical hyperalgesia tests, respectively. The animals were euthanized for analysis of cannabinoid receptor type 1 (CB1) and type 2 (CB2), and µ-opioid receptor (MOR) in the anterior cingulate cortex (ACC), periaqueductal gray matter (PAG), and thalamus areas by immunohistochemistry (IHC) and Western blotting. Our data revealed a decrease in the nociceptive threshold in both forepaws after surgery; in contrast, there was improvement in painful symptoms after the exercise protocol. For cannabinoid system there were an increase in CB2 expression in the ACC and PAG, and in CB1 levels in the PAG. And for opioid system there was an increase of MOR expression in the thalamus. Thus, modulation of those receptors by physical exercise can be an important non-pharmacological intervention to reduce painful symptoms in a rat model of PD, contributing to knowledge and promotion of better treatment aimed at improving the quality of life of PD patients.
Assuntos
Nociceptividade/fisiologia , Doença de Parkinson/metabolismo , Doença de Parkinson/psicologia , Condicionamento Físico Animal , Receptor CB1 de Canabinoide/metabolismo , Receptor CB2 de Canabinoide/metabolismo , Receptores Opioides mu/metabolismo , Animais , Modelos Animais de Doenças , Giro do Cíngulo/metabolismo , Hiperalgesia/complicações , Hiperalgesia/prevenção & controle , Doença de Parkinson/prevenção & controle , Substância Cinzenta Periaquedutal/metabolismo , Ratos WistarRESUMO
Objetivou-se estudar o efeito do ômega 3 e da vitamina B12 no espermograma, na histomorfometria dos órgãos reprodutivos e na temperaturas do corpo com termografia infravermelha em ratos Wistar. Utilizaram-se 16 ratos, em quatro grupos (n=4), que receberam injeções diárias por 30 dias, sendo: grupo controle - solução salina; grupo ômega 3 - óleo de peixe 1g/kg; grupo B12 - vitamina B12 3µg; e grupo ômega 3 + B12 - óleo de peixe 1g/kg e vitamina B12 3µg. Imagens termográficas de áreas do corpo foram obtidas. No 30º dia, os ratos foram sacrificados e realizaram-se as análises de morfologia espermática e histomorfometria. Os dados foram submetidos à análise de variância e ao teste de Tukey a 5%. A temperatura da superfície do escroto foi superior no grupo B12 (P<0,05). Não houve diferenças entre grupos (P>0,05) para temperaturas do globo ocular. Houve correlação entre temperatura da superfície do escroto e porcentagem de gota citoplasmática distal (P=0,678). A elevação da temperatura do escroto resulta no aumento da porcentagem de gotas citoplasmáticas distais. A temperatura do globo ocular não sofre influência significativa do ômega 3 e da vitamina B12. O ômega 3 reduz o epitélio seminífero, e a vitamina B12 minimiza esse efeito.(AU)
The objective of this study was to study the effect of Omega 3 and vitamin B12 on spermogram, histomorphometry of reproductive organs and body temperature with infrared thermography in Wistar rats. Sixteen rats were used in four groups (n= 4) who received daily injections for 30 days. Control Group - saline solution; Group Omega 3 - fish oil 1g/kg; Group B12 - vitamin B12 3μg and Group Omega 3 + B12 - fish oil 1g/kg and vitamin B12 3μg. Thermographic images of body were obtained. On the 30th day the rats were sacrificed and analyzes of sperm morphology and histomorphometry were performed. Data were submitted to analysis of variance and Tukey's test at 5%. The surface temperature of the scrotum was higher in group B12 (P< 0.05). There were no differences between groups (P> 0.05) for eyeball temperatures. There was a correlation between scrotal temperature and distal cytoplasmic droplet (P= 0.678). Elevation of scrotum temperature results in an increase in the percentage of distal cytoplasmic droplets. The temperature of the eyeball is not significantly influenced by Omega 3 and vitamin B12. Omega 3 reduces the seminiferous epithelium and vitamin B12 minimizes this effect.(AU)
Assuntos
Animais , Ratos , Vitamina B 12/efeitos adversos , Ácidos Graxos Ômega-3/efeitos adversos , Ratos Wistar/metabolismo , Contagem de Espermatozoides/veterinária , Termografia/veterináriaRESUMO
O conhecimento do desenvolvimento mamário de um rebanho leiteiro é fundamental, pois relaciona-se à eficiência produtiva. Objetivou-se avaliar a associação dos parâmetros térmicos mamários com concentrações hormonais de búfalas em distintos estágios fisiológicos. Foram utilizadas 24 búfalas mestiças Murrah, em quatro grupos (n= 6): grupo 1 (bezerras), grupo 2 (novilhas), grupo 3 (gestantes) e grupo 4 (lactantes). A cada 28 dias, durante 4 meses, realizaram-se exames de termografia digital por infravermelho para verificar temperatura superficial dos corpos mamários craniais (CMCr) e caudais (CMC), das cisternas craniais (CGMCr) e caudais (CGMC) e tetas craniais (TGMCr) e caudais (TGMC). Foi aferida temperatura retal (TR) e colhido sangue para mensuração das concentrações plasmáticas do fator semelhante a insulina tipo-I, insulina (INS), hormônio do crescimento (GH), progesterona (P4) e estradiol. Grupos 1 e 2 apresentaram correlação de TR com CGMCr. No grupo 3, TR correlacionou-se com TGMCr, TGMC e concentrações plasmáticas de P4. No grupo 4, houve correlação de TR com CGMC, TGMCr e concentrações plasmáticas de INS e GH, e de TGMC com concentrações plasmáticas de GH. Nos quatro grupos, CGMCr correlacionou-se com CMCr e TGMCr, semelhantemente ao observado na porção caudal. Variações térmicas mamárias refletiram alterações fisiológicas aguardadas nos períodos avaliados.(AU)
The knowledge of the mammary development of a dairy herd is key, since it is related to its productive efficiency. The objective of this study was to evaluate the association of mammary thermal parameters with hormonal concentrations of buffaloes at different physiological stages. Twenty-four Murrah crossbred buffaloes were used in four groups (n= 6): group 1 (calves), group 2 (heifers), group 3 (pregnant animals) and group 4 (lactating animals). Every 28 days, for 4 months, infrared digital thermography was performed to check the surface temperature of cranial (CrCM) and caudal (CCM) corpus mammae, cranial (CrC) and caudal (CC) cisterns and cranial (CrT) and caudal teats (CT). Rectal temperature (RT) was measured and blood was collected to measure plasma concentrations of insulin-like factor I, insulin (INS), growth hormone (GH), progesterone (P4) and estradiol. Groups 1 and 2 presented correlation of RT with CrC. In group 3, RT correlated with CrT, CT and plasma P4 concentrations. In group 4, there was correlation of RT with CC, CrT and plasma concentrations of INS and GH, and CT with plasma GH concentrations. In all four groups, CrC correlated with CrCM and CrT, similar to that observed in the caudal portion. The thermal changes in the mammary glands reflected the expected physiological changes in the evaluated periods.(AU)
Assuntos
Animais , Feminino , Bovinos , Bovinos/fisiologia , Glândulas Mamárias Animais/crescimento & desenvolvimento , Termografia/classificação , Búfalos/fisiologia , Técnicas de Diagnóstico Endócrino/veterináriaRESUMO
The aim was to study pregnant cervus in captivity, to obtain body morphometric data, serum progesterone concentrations, and pH of the vagina. Females of cervus (Cervus unicolor), (n=8) between 2 and 3 years old were used in November during breeding season. After sedation, in one collection, the following was measured: body weight 54.50 ± 18.70kg, body length 1.21 ± 0.16m, height at withers 0.75 ± 0.08m, thoracic perimeter 0.84 ± 0.12m, and body mass index 97.32 ± 10.50kg / m2. After sedation, blood samples were collected to measure progesterone concentration by radioimmunoassay and the use of an indicator tape to obtain the pH of the vagina was performed eight times at four day intervals in the eight females, in a total of 64 samples for each exam. Data were analyzed, and significance was at 5%. There were no differences (P> 0.05) between measurements for serum progesterone concentrations and pH of the vagina. The correlations between body weight, progesterone, pH, and body mass index were not significant (P> 0.05). There was no significant oscillation of progesterone concentration and pH of the vagina in the pregnancy period studied.(AU)
Assuntos
Animais , Proteínas Sanguíneas/análise , Cervos/anatomia & histologia , Progesterona/análise , Endocrinologia , Concentração de Íons de HidrogênioRESUMO
The aim was to study pregnant cervus in captivity, to obtain body morphometric data, serum progesterone concentrations, and pH of the vagina. Females of cervus (Cervus unicolor), (n=8) between 2 and 3 years old were used in November during breeding season. After sedation, in one collection, the following was measured: body weight 54.50 ± 18.70kg, body length 1.21 ± 0.16m, height at withers 0.75 ± 0.08m, thoracic perimeter 0.84 ± 0.12m, and body mass index 97.32 ± 10.50kg / m2. After sedation, blood samples were collected to measure progesterone concentration by radioimmunoassay and the use of an indicator tape to obtain the pH of the vagina was performed eight times at four day intervals in the eight females, in a total of 64 samples for each exam. Data were analyzed, and significance was at 5%. There were no differences (P> 0.05) between measurements for serum progesterone concentrations and pH of the vagina. The correlations between body weight, progesterone, pH, and body mass index were not significant (P> 0.05). There was no significant oscillation of progesterone concentration and pH of the vagina in the pregnancy period studied.
Assuntos
Animais , Cervos/anatomia & histologia , Progesterona/análise , Proteínas Sanguíneas/análise , Concentração de Íons de Hidrogênio , EndocrinologiaRESUMO
Chronic constriction injury (CCI) simulates the symptoms of chronic nerve compression, which is characterized by allodynia and hyperalgesia. Nerve growth factor (NGF) is released after nerve injury by immune and Schwann cells and transported in retrograde fashion to the dorsal root ganglion (DRG), resulting in increased synthesis of Substance P (SP) and the triggering of neuropathic pain. Here we performed long-term evaluation of allodynia and hyperalgesia in a CCI model, and evaluated the effects of NGF and SP on the peripheral and central nervous systems. Most previous studies have shown deficits and molecular changes 14 days after surgery, however, the long-term effects have not been evaluated. We performed Randall-Selitto, Von Frey, Hargreaves and acetone tests for the entire 56 days post-surgery. Several of these deficits increased 14 to 56 days after CCI and we measured a constant increase in NGF levels in the DRG and spinal cord over the course of the experiment. In contrast, SP optical density maintained enhanced expression in DRG tissue from 14 to 56 days after CCI, whereas it was significantly increased only 56 days post-surgery in spinal cord. We perform long-term evaluation of symptoms associated with CCI and measure associated molecular changes. Moreover, by characterizing the behavioral signatures of this model, our work supports future studies.
Assuntos
Hiperalgesia/metabolismo , Fator de Crescimento Neural/metabolismo , Nervo Isquiático/lesões , Nervo Isquiático/metabolismo , Substância P/metabolismo , Animais , Doença Crônica , Gânglios Espinais/metabolismo , Gânglios Espinais/patologia , Gânglios Espinais/fisiopatologia , Hiperalgesia/patologia , Hiperalgesia/fisiopatologia , Masculino , Ratos , Ratos Wistar , Nervo Isquiático/patologia , Nervo Isquiático/fisiopatologia , Medula Espinal/metabolismo , Medula Espinal/patologia , Medula Espinal/fisiopatologiaRESUMO
Brucellosis remains as a major infectious disease of domestic animals and is considered a re-emerging zoonosis in several countries. B. abortus infections in bulls are related to reproductive tract infections, although infected animals show transient serological titers or nonreactor status. Thus, diagnosis of bovine brucellosis based exclusively on serological tests probably underestimates B. abortus infections in bulls. In this scenario, three hundred thirty-five serum samples from reproductively mature bovine bulls were subjected simultaneously to standard serodiagnosis using the rose Bengal test (RBT), 2-mercaptoethanol (2-ME), complement fixation (CFT), and fluorescence polarization assay (FPA). Furthermore, conventional semen plasma agglutination (SPA) and modified 2-ME, FC and, FPA were carried out in all bulls replaing serum by seminal plasma. Semen from all bulls was also analyzed for sperm viability, microbiological culture in Farrell media, and polymerase chain reaction (PCR). Only eight (2.38%) semen samples were considered improper for reproduction services (necrospermia and azoospermia), although none of these animals was positive in any of the diagnosis methods used. Five bulls (1.49%) were simultaneously positive in conventional RBT, 2-ME, SPA, modified 2-ME, microbiological culture in Farrell media, and in PCR for B. abortus strain 19. Two (1.67%) bulls were positive in PCR for B. abortus field strains and negative in all other tests, although semen was considered viable to reproduction service. The identification of B. abortus B19 strain in serum and semen of bulls occurred probably due to improper vaccination of males or infection by B19 strain shedding by vaccinated females that could to contaminated environment of farms. In addition, detection of B. abortus field strains only using PCR in bulls without sperm viability abnormalities indicate the need for including molecular methods to improve diagnosis of the disease in bovine bulls.
Assuntos
Brucella abortus , Brucelose Bovina/sangue , Brucelose Bovina/microbiologia , Sêmen/microbiologia , Animais , Bovinos , MasculinoRESUMO
Orofacial pain is associated with peripheral and central sensitization of trigeminal nociceptive neurons. Nerve injury results in release of chemical mediators that contribute to persistent pain conditions. The activation of the transient receptor potential vanilloid 1 (TRPV1), promotes release of calcitonin gene-related peptide (CGRP) and substance P (SP) from trigeminal nerve terminals. CGRP and SP contribute to the development of peripheral hyperalgesia. The expression of SP and CGRP by primary afferent neurons is rapidly increased in response to peripheral inflammation. CGRP receptor activation promotes activation of AMPA receptors, leading to increased firing of neurons which is reflected as central sensitization. In this study we investigated whether inferior alveolar nerve (IAN) injury influences AMPA receptors, CGRP, SP and TRPV1 expression in the trigeminal ganglion (TG). The relative expression of the protein of interest from naive rats was compared to those from injured rats and animals that received low level laser therapy (LLLT). IAN-injury did not change expression of GluA1, GluA2 and CGRP, but increased the expression of TRPV1 and SP. LLLT increases GluA1 and GluA2 expression and decreases TVPV1, SP and CGRP. These results, together with previous behavioral data, suggest that IAN-injury induced changes in the proteins analyzed, which could impact on nociceptive threshold. These data may help to understand the molecular mechanisms of pain sensitization in the TG.
Assuntos
Traumatismos do Nervo Facial/radioterapia , Regulação da Expressão Gênica/efeitos da radiação , Terapia com Luz de Baixa Intensidade , Nervo Mandibular/efeitos da radiação , Gânglio Trigeminal/efeitos da radiação , Animais , Peptídeo Relacionado com Gene de Calcitonina/genética , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Traumatismos do Nervo Facial/genética , Traumatismos do Nervo Facial/metabolismo , Traumatismos do Nervo Facial/patologia , Masculino , Nervo Mandibular/metabolismo , Nervo Mandibular/patologia , Neurônios Aferentes/metabolismo , Neurônios Aferentes/patologia , Neurônios Aferentes/efeitos da radiação , Estimulação Luminosa/métodos , Ratos , Ratos Sprague-Dawley , Receptores de AMPA/genética , Receptores de AMPA/metabolismo , Transdução de Sinais , Substância P/genética , Substância P/metabolismo , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismo , Gânglio Trigeminal/lesões , Gânglio Trigeminal/metabolismoRESUMO
In the present work, we investigated the antinociceptive effect of gabapentin in a chronic myositis model and its interference in spinal glial cells. Chronic myositis was induced by injection of Complete Freund Adjuvant (CFA) into the right gastrocnemius (GS) muscle of rats and tests for evaluating mechanical hyperalgesia, thermal hyperalgesia and tactile allodynia were performed. Pharmacological treatment with gabapentin was administrated intrathecally and 100µg and 200µg doses were tested. For analyzing astrocytes and microglia in the spinal cord, immunochemistry assay was performed. It was found that gabapentin 200µg reverted CFA-induced chronic muscle pain bilaterally, in all applied tests and it was able to attenuate microglial but not astrocytes activation in the dorsal horn of spinal cord. In conclusion, gabapentin was able to inhibit hyperalgesia and allodynia in chronic myositis and also to attenuate spinal microglial activation. Therefore, gabapentin could be used as treatment for targeting chronic muscle pain.
Assuntos
Aminas/farmacologia , Ácidos Cicloexanocarboxílicos/farmacologia , Hiperalgesia/tratamento farmacológico , Hiperalgesia/patologia , Microglia/efeitos dos fármacos , Microglia/patologia , Miosite/complicações , Ácido gama-Aminobutírico/farmacologia , Aminas/uso terapêutico , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/patologia , Comportamento Animal/efeitos dos fármacos , Contagem de Células , Doença Crônica , Ácidos Cicloexanocarboxílicos/uso terapêutico , Gabapentina , Hiperalgesia/complicações , Masculino , Mialgia/complicações , Ratos , Ratos Wistar , Corno Dorsal da Medula Espinal/patologia , Ácido gama-Aminobutírico/uso terapêuticoRESUMO
BACKGROUND: Nerve injury often results in persistent or chronic neuropathic pain characterized by spontaneous burning pain accompanied by allodynia and hyperalgesia. Low level laser therapy (LLLT) is a noninvasive method that has proved to be clinically effective in reducing pain sensitivity and consequently in improving the quality of life. Here we examined the effects of LLLT on pain sensitivity induced by chronic constriction injury (CCI) in rats. CCI was performed on adult male rats, subjected thereafter to 10 sessions of LLLT, every other day, and starting 14 days after CCI. Over the treatment period, the animals were evaluated for nociception using behavioral tests, such as allodynia, thermal and mechanical hyperalgesia. Following the sessions, we observed the involvement of satellite glial cells in the dorsal root ganglion (DRG) using immunoblotting and immunofluorescence approaches. In addition we analyzed the expression levels of interleukin 1 (IL-1ß) and fractalkine (FKN) after the same stimulus. RESULTS: LLLT induced an early reduction (starting at the second session; p ≤ 0.001) of the mechanical and thermal hyperalgesia and allodynia in CCI rats, which persisted until the last session. Regarding cellular changes, we observed a decrease of GFAP (50%; p ≤ 0.001) expression after LLLT in the ipsilateral DRG when compared with the naive group. We also observed a significant increase of pro-inflammatory cytokines after CCI, whereas LLLT dramatically inhibited the overexpression of these proteins. CONCLUSIONS: These data provide evidence that LLLT reverses CCI-induced behavioral hypersensitivity, reduces glial cell activation in the DRG and decreases pro-inflammatory cytokines; we suggest that this involvement of glial cells can be one potential mechanism in such an effect.
Assuntos
Lasers Semicondutores/uso terapêutico , Terapia com Luz de Baixa Intensidade , Neuralgia/radioterapia , Animais , Quimiocina CX3CL1 , Ensaio de Imunoadsorção Enzimática , Gânglios Espinais/metabolismo , Gânglios Espinais/patologia , Hiperalgesia/patologia , Hiperalgesia/radioterapia , Interleucina-1/análise , Masculino , Microscopia de Fluorescência , Neuralgia/patologia , Neuroglia/citologia , Neuroglia/metabolismo , Neuroglia/efeitos da radiação , Ratos , Ratos WistarRESUMO
A coloração pela prata das regiões organizadoras de nucléolos (NORs) é caracterizada por marcar proteínas ligadas ao ácido ribonucleico ribossômico, avaliando a proliferação em células normais ou neoplásicas. Objetivou-se estudar, em testículos de ovinos obtidos em matadouro, a validade do uso da técnica de coloração pela prata (AgNOR) na identificação das regiões organizadoras de nucléolo (NORs) em células saudáveis da linhagem espermatogênica. Utilizaram-se 43 pares de testículos de ovinos mestiços entre seis e 10 meses de idade. Testes de Wilcoxon e Spearman foram empregados, com nível de 5%. As médias das NORs nas células das gônadas direita e esquerda foram, respectivamente: espermatogônia (8,77±1,14 e 9,04±0,96), espermatócitos (4,99±2,00 e 6,20±2,07; P<0,05), Leydig (8,05±2,82 e 7,89±2,29) e Sertoli (8,07±1,88 e 7,61±2,16; P<0,05). Houve correlação (P<0,05) entre os lados para o número de NORs: espermatócitos x Leydig (0,49); espermatócitos x Sertoli (0,49) e Leydig x Sertoli (0,96). Conclui-se ser válido o emprego da técnica AgNOR para avaliar o potencial proliferativo das células saudáveis em testículos de ovinos com prática execução e baixo custo.(AU)
The silver staining technique for AgNOR nucleolar organizer regions (NORs) is characterized by marking proteins linked to the ribosomal ribonucleic acid, evaluating cell proliferation. The aim was to study the validity of the AgNOR staining technique in the testicular cell proliferation in crossbred ovine. Forty-three pairs of ovine testicles between 6 and 10 months old were collected. Wilcoxon and Spearman tests were used with a significance level of 5%. The mean NORs count in cells of the right and left gonads were respectively: spermatogonia (8.77±1.14 and 9.04±0.96), spermatocytes (4.99±2.00 and 6.20±2.07, P<0.05), Leydig (8.05±2.82 and 7.89±2.29) and Sertoli cells (8.07±1.88 and 7.61±2.16; P<0.05). There was a correlation between the mean values for the right and left sides for the number of NORs (P<0.05) between Leydig x spermatocytes (0.49); spermatocytes x Sertoli (0.49) and Sertoli x Leydig (0.96). The study demonstrates that the AgNOR staining technique is indicated to evaluate the cell proliferative potential in ovine testis with practical implementation and low cost.(AU)
Assuntos
Animais , Masculino , Testículo , Ovinos , Nucléolo Celular/ultraestrutura , Coloração pela Prata/veterinária , Proliferação de CélulasRESUMO
Objetivou-se estudar a morfometria corpórea, as características do sêmen, o perfil proteico do plasma seminal em SDS-PAGE e a concentração sérica de testosterona em cervos-sambar (Cervus unicolor), criados em cativeiro, na estação reprodutiva da primavera. Quatro machos com idades entre 12 e 36 meses foram avaliados em quatro momentos, com intervalos de sete dias, com peso corpóreo (60,5 a 89,0kg), índice de massa corporal (93,07kg/m2 a 126,56kg/m2), volume do ejaculado (0,50±0,35mL a 0,75±0,28mL), motilidade espermática (87,75±4,78% a 90,00±7,07%), defeitos totais (17,25±5,81% a 47,72±17,55%), testosterona sérica (6,43±4,33ng/dL a 166,00±64,48ng/dL) e proteínas do plasma seminal com bandas entre 7,6 e 142kDa. As características dos ejaculados não diferiram (P>0,05) entre as três primeiras colheitas. Houve diferença (P<0,05) para os defeitos espermáticos com elevação na quarta colheita. No plasma seminal de cada cervo, foram identificadas de 16 a 27 bandas de proteínas entre 7,6 e 142kDa. Conclui-se que a qualidade espermática foi satisfatória na primavera. O estresse das contenções sucessivas causou queda da qualidade espermática. A idade influi na concentração sérica de testosterona, a qual foi maior nos cervos aos 36 meses...
The aim of this work was to study the body morphometry, semen characteristics, seminal plasma protein profile in SDS-PAGE and serum testosterone concentration in Sambar Deer (Cervus unicolor), in captivity in the breeding season (spring). Four males aged between 12 and 36 months were assessed in four moments with intervals of seven days with body weight (60.5 to 89.0kg), body mass index (93.07 to 126.56kg/m2), ejaculate volume (0.50±0.35mL to 0.75±0.28mL), sperm motility (87.75±4.78% to 90.00±7.07% ), total defects (17.25±5.81% to 47.72±17.55%), serum testosterone (6.43±4.33 ng/dL to 166.00±64.48ng/dL) and seminal plasma proteins with bands between 7.6 and 142 kDa. The characteristics of ejaculates did not differ (P>0.05) among ejaculates (1st, 2nd and 3rd). There were differences (P<0.05) for sperm defects elevation on the fourth ejaculate. In seminal plasma 16 to 27 protein bands between 7.6 and 142 kDa were identified. In conclusion, sperm quality was satisfactory in the spring and the stress of successive contentions decreased sperm quality. Also, there is influence of age upon serum testosterone concentration which was higher in deer at 36 months...
Assuntos
Animais , Análise do Sêmen/veterinária , Cervos , Capacitação Espermática , Androgênios/análise , Eletroforese em Gel de Poliacrilamida/veterinária , Testosterona/análiseRESUMO
Estudou-se a influência das quatro estações do ano nas características do sêmen e nas concentrações de testosterona e cortisol em touros. Cinco touros Nelore e cinco Simental entre 48 e 72 meses de idade, criados extensivamente, foram avaliados andrologicamente por meio de exames físicos das características morfológicas do sêmen e das concentrações séricas de testosterona e cortisol. Houve redução na motilidade e no vigor do sêmen no inverno (P<0,05) na raça Simental. Observou-se correlação (P<0,01) entre testosterona x motilidade (0,69) e testosterona x vigor (0,57) na raça Simental, e cortisol x motilidade (0,68) e cortisol x vigor (0,65) na raça Nelore. O efeito das estações do ano modificou a qualidade do sêmen com aumento da motilidade e vigor espermáticos na primavera e verão nos touros Simental. A concentração de cortisol diminuiu no outono nos touros Nelore.
The influence of four seasons per year on semen characteristics and levels of testosterone and cortisol in Nelore and Simental bulls was studied. Five Nelore and five Simental bulls extensively managed, at 48-72 months old were evaluated for sexual soundness using physical and morphological characteristics of semen and serum levels of testosterone and cortisol. In Simental bulls there was decreased motility and vigor of semen (P<0.05) during winter. There was a correlation (P<0.01) between testosterone x motility (0.69) and testosterone x vigor (0.57) in the Simental breed and cortisol x motility (0.68) and cortisol x vigor (0.65) in Nelore the breed. The effect of seasons during the year changed the semen quality with increased sperm motility and vigor in spring-summe r in Simental bulls. The cortisol level decreased in autumn in Nelore bulls.
RESUMO
Nerve injury leads to a neuropathic pain state that results from central sensitization. This phenomenom is mediated by NMDA receptors and may involve the production of nitric oxide (NO). In this study, we investigated the expression of the neuronal isoform of NO synthase (nNOS) in the spinal cord of 3-month-old male, Wistar rats after sciatic nerve transection (SNT). Our attention was focused on the dorsal part of L3-L5 segments receiving sensory inputs from the sciatic nerve. SNT resulted in the development of neuropathic pain symptoms confirmed by evaluating mechanical hyperalgesia (Randall and Selitto test) and allodynia (von Frey hair test). Control animals did not present any alteration (sham-animals). The selective inhibitor of nNOS, 7-nitroindazole (0.2 and 2 microg in 50 microL), blocked hyperalgesia and allodynia induced by SNT. Immunohistochemical analysis showed that nNOS was increased (48% by day 30) in the lumbar spinal cord after SNT. This increase was observed near the central canal (Rexed's lamina X) and also in lamina I-IV of the dorsal horn. Real-time PCR results indicated an increase of nNOS mRNA detected from 1 to 30 days after SNT, with the highest increase observed 1 day after injury (1469%). Immunoblotting confirmed the increase of nNOS in the spinal cord between 1 and 15 days post-lesion (20%), reaching the greatest increase (60%) 30 days after surgery. The present findings demonstrate an increase of nNOS after peripheral nerve injury that may contribute to the increase of NO production observed after peripheral neuropathy.