ABSTRACT
ABSTRACT BACKGROUND: Few studies regarding arthritic diseases have been performed to verify the presence of the neurodegeneration. Given the increased oxidative stress and extra-articular effects of the rheumatoid arthritis, the gastrointestinal studies should be further investigated aiming a better understanding of the systemic effects the disease on enteric nervous system. OBJECTIVE: To determine whether the rheumatoid arthritis affects the nitrergic density and somatic area of the nNOS- immunoreactive (IR) myenteric neurons, as well as the morphometric areas of CGRP and VIP-IR varicosities of the ileum of arthritic rats. METHODS: Twenty 58-day-old male Holtzmann rats were distributed in two groups: control and arthritic. The arthritic group received a single injection of the Freund's Complete Adjuvant in order to induce arthritis model. The whole-mount preparations of ileum were processed for immunohistochemistry to VIP, CGRP and nNOS. Quantification was used for the nitrergic neurons and morphometric analyses were performed for the three markers. RESULTS: The arthritic disease induced a reduction 6% in ileal area compared to control group. No significant differences were observed in nitrergic density comparing both groups. However, arthritic group yielded a reduction of the nitrergic neuronal somatic area and VIP-IR varicosity areas. However, an increase of varicosity CGRP-IR areas was also observed. CONCLUSION: Despite arthritis resulted in no alterations in the number of nitrergic neurons, the retraction of ileal area and reduction of nitrergic somatic and VIP-IR varicosity areas may suggest a negative impact the disease on the ENS.
RESUMO CONTEXTO: Poucos estudos sobre doenças artríticas têm sido realizados para verificar a presença de neurodegeneração. Diante do aumento do estresse oxidativo e dos efeitos extra-articulares da artrite reumatoide, estudos gastrointestinais devem ser investigados visando uma melhor compreensão dos efeitos sistêmicos da doença no sistema nervoso entérico. OBJETIVO: Determinar se a artrite reumatoide afeta a densidade nitrérgica e a área somática dos neurônios mioentéricos imunorreativos ao nNOS (nNOS-IR), bem como para as áreas morfométricas das varicosidades CGRP-IR e VIP-IR do íleo de ratos artríticos. MÉTODOS: Vinte ratos Holtzmann, com 58 dias de idade, foram distribuídos em dois grupos: controle e artrítico. O grupo artrítico recebeu uma única injeção do adjuvante completo de Freund para induzir o modelo de artrite. Os preparados totais de íleo foram processados para imuno-histoquímica ao VIP, CGRP e nNOS. A quantificação foi utilizada para os neurônios nitrérgicos e as análises morfométricas foram realizadas para os três marcadores. RESULTADOS: A doença artrítica induziu uma redução de 6% na área ileal em relação ao grupo controle. Não foram observadas diferenças significativas na densidade nitrérgica comparando os dois grupos. No entanto, o grupo artrítico produziu uma redução da área somática neuronal nitrérgica e da área das varicosidades do VIP-IR. Entretanto, foi observado um aumento das áreas das viricosidades CGRP-IR. CONCLUSÃO: Apesar da artrite não resultar em alterações no número de neurônios nitrérgicos, a retração da área ileal e a redução das áreas somática nitrérgica e das varicosidades do VIP-IR podem sugerir um impacto negativo da doença no sistema nervoso entérico.
Subject(s)
Animals , Male , Rats , Arthritis, Rheumatoid/physiopathology , Enteric Nervous System/physiopathology , Nitrergic Neurons/physiology , Nitric Oxide Synthase Type I/metabolism , Immunohistochemistry , Rats, Sprague-Dawley , Nitrergic Neurons/metabolism , Disease Models, Animal , Nitric Oxide Synthase Type I/physiology , Myenteric Plexus/physiopathology , Myenteric Plexus/metabolismABSTRACT
Avaliaram-se os efeitos crônicos de uma dieta severamente hipoprotéica, sobre aspectos quantitativos e morfométricos de neurônios mientéricos, NADPH-diaforase positivos do ceco de ratos jovens. Oito ratos foram divididos em dois grupos, sendo que para um dos grupos foi oferecida ração com teor protéico de 26% (controle) e, para o outro, ração com o teor protéico de 4%, mantendo-se o balanço vitamínico e mineral durante 12 semanas. Após eutanásia, o ceco de cada animal foi coletado e dissecado para confecção de preparados de membrana, os quais foram submetidos à técnica da NADPH-diaforase. Para contagem e mensuração destes neurônios, o ceco foi dividido em quatro regiões: apical mesentérica; apical antimesentérica; basal antimesentérica; próxima à ampola cecal. Não se observou alteração significativa para o peso dos animais, área do ceco e densidade populacional dos neurônios mientéricos no final do experimento. Por outro lado, em algumas regiões do ceco, os neurônios tiveram redução da área nuclear, com conseqüente redução da área do pericário
The chronicle effects of a severe hypoproteic diet on quantitative and morphometric aspects of NADPHdiaphorase-positive myenteric neurons from the cecum of young rats were assessed. Eight rats were divided into two groups. One group (control) was fed with 26%-protein chow and the other with 4%-protein chow during 12 weeks. The vitamin and mineral balance was kept. After euthanasia, the cecum of each animal was collected and dissected for the whole-mount preparation, which were submitted to NAPDH-diaphorase. The cecum was divided into four regions for counting and measurement of neurons: apical mesenteric, antimesenteric apical, basal antimesenteric, and next to the cecal ampulla. Significant alterations were not noted with respect to the weight of the animals, cecum area, and population density of myenteric neurons at the end of the experiment. On the other hand, in some regions of the cecum, the neurons presented a reduction of the nuclear area with resulting reduction of the soma area
Se evaluaron los efectos crónicos de una dieta severamente hipoprotéica, sobre aspectos cuantitativos y morfométricos de neuronas mientéricos, NADPH-diaforase positivos del ceco de ratas jóvenes. Ocho ratas fueron divididas en dos grupos, siendo que para uno de los grupos fue ofrecida ración con tenor proteico de 26% (control) y, para el otro, ración con el tenor proteico de 4%, manteniéndose el balance vitamínico y mineral durante 12 semanas. Tras eutanasia, El ceco de cada animal fue recolectado y disecado para confección de preparados de membrana, los cuales fueron sometidos a la técnica de la NADPH-diaforase. Para recuento y mensuración de estas neuronas, el ceco fue dividido en cuatro regiones: apical mesentérica, apical antimesentérica, basal antimesentérica y próxima a la ampolla cecal. No se observó alteración significativa en el peso de los animales, área del ceco y densidad poblacional de las neuronas mientéricas al final Del experimento. Por otro lado, en algunas regiones del ceco las neuronas tuvieron reducción del área nuclear, con consecuente reducción del área del pericario
Subject(s)
Animals , Protein-Energy Malnutrition/veterinary , Neurons , Myenteric Plexus/anatomy & histology , RatsABSTRACT
In order to characterize the neurochemical features of nitrergic myenteric neurons in the rat esophagus and to disclose some possible cause of nitrergic neuronal cell loss in the rat esophagus during aging, whole-mount preparations from different esophageal segments of different age- and strain-groups were studied by double immunohistochemistry of neuronal nitric oxide synthase (nNOS) and calbindin (CB). We found for the first time co-localization of nNOS and CB in myenteric neuronal bodies with a specific distribution pattern along the entire length of rat esophagus: significantly higher percentage of nNOS/CB immunoreactive (ir) myenteric neurons (ca 30%-40%) were found in the abdominal segment in comparison with the other segments of the organ (P<0.001). Nitrergic myenteric neurons in the rat esophagus could be subdivided at least into two groups: nNOS+/CB+ and nNOS+/CB-. Moreover, the majority of CB-ir myenteric neurons,especially in the abdominal esophagus expressed nNOS-ir (ca 95%-100%). All these features were persisted in all age and strain groups of rats studied. The relative percentages of the nNOS-/CB-ir neurons among the total nitrergic population varied strain-dependently during aging. Regression analysis yielded a significant negative correlation between the percentage of nNOS-/CB-ir neurons found in the young Sprague-Dawley rats and percentage of nitrergic neuronal loss during aging (correlation coefficient 0.99; P< 0.05). However, the functional significance of the regional differences in nNOS-/CB-ir neurons along the rat esophagus remains to be elucidated.
ABSTRACT
Objective To observe the changes in enteric motor function and acetylcholine esterase(AchE)neuron and nitric oxide synthase neuron(NOS)in enteric nervous system after different abdominal operations,and to explore the mechanism of these changes.Methods Sprague Dawley rats were divided into experimental groups(laparotomy group,intestine massage group and intestinal operation group)and the control group.Rats in each group were gavaged with medicinal carbon powder,and then the transimission of carbon powder in small intestine was determined,and the spreading specimens of intestinal myenteric plexus of small intestine and large intestine were collected and stained with AchE and nicotinamide-adenine dinucleotide phosphate-D(NADPH-d)for histological study,and the density of distribution and positive stained AchE and NOS neurons were observed and compared.Results The changes in enteric motor function:the propellant rate of carbon powder was slower in enteric massage group and intestinal operation group than that in laparotomy group and control group(18.1?2.8,25.7?3.5 vs 47.4?3.6,49.8?4.5,P0.05).Compared with the control group,the number and positive expression of AchE positive neuron in intestinal myenteric plexus decreased in small intestine and large intestine in intestinal operation group and intestine massage group(small intestine 43.0?3.0,52.2?3.3 vs 65.3?3.9,P
ABSTRACT
Objective To explore the development of nitrergic neurons in the tongue of human fetus. Methods Using histochemical method of NADPH\|diaphorase, the differentiation, migration and development of the nitrergic neurons in the tongue of human fetus were studied. Results At the fourth month of the gestation, the round cells of epithelial tissue in tongue differentiated into fusiform nitrergic nerve cells, which moved from the epithelial tissue to subepithelial and muscular tissue. The bodies of nitrergic neurons were small, the positive reaction of nitric oxide synthase(NOS) was weak. The development of the process of nitrergic neurons might be divided into two phases:The first phase, the growth and development phase, occured from the fourth to the seventh month, the somatic size enlarged gradually, the numbers increased, the positive reaction of NOS increased gradually and reached its peak at the seventh month. The morphology of neurons was characterized by the development from the fusiform form to the tadpole form and then a diversity of shape. The second phase, the maturation phase, denoted the eighth to tenth month, the bodies of nitrergic neurons were obviously enlarged, the staining intensity of NOS was increased. The nitrergic neurons were scattered in the subepithelial layer and muscular tissue and the typical ganglionic cells were observed. Conclusion Nitrergic neurons of tongue originate from its epithelial tissue at the time of early fetal development. By the differentiation, migration, multiplication, growth and maturation, mature nitrergic neurons are formed. [