Morphometry and quantification of the myenteric neurons of the cecum of rats (rattus norvegicus) submitted to experimental chronic alcoholism / Morfometría y cuantificación de las neuronas mientéricas del ciego en ratones (rattus norvegicus) sometidos a alcoholismo crónico experimental

Morphometric and quantitative analyses were accomplished to study the effects of the consumption of alcohol above the size and density of the myenteric neurons of cecum of rats. Ten rats with 90 days were divided in the groups: control (C), and alcoholic (A). After 120 days of treatment with ethanol the cecum of both groups were collected, submitted it prepared of membranes that after carried out to Giemsa´s technique, permitted to evaluate the neuronal density, in an area of 13.44mm², and measure the cell body area of 300 neurons by group. The alcoholic rats presented an increase in the number of small neurons and a reduction of the big and medium neurons. The neuronal density verified in alcoholic rats was significantly reduced regarding the controls rats, however, that reduction left of be statistically significant when was projected the neuronal density for the total area of cecum, since the macroscopic observation showed that the alcoholic rats presented a cecum dilated. The alcoholism induced a significant reduction in the final body weight of the rats of the GA, provoked enlargement of cecum of the rats causing to a big dispersion neuronal. The enlargement of cecum of the alcoholic rats is probably associated with the functional alterations of the myenteric neurons that have repercussions in the tone of the intestinal smooth muscle.

Los análisis morfométrico y cuantitativo fueron hechos para estudiar los efectos del consumo de alcohol sobre el tamaño y densidad de las neuronas mientéricas del ciego de ratones. Diez ratones de 90 días, fueron divididos en los grupos: control (C), y alcohólico (A). Después de 120 días de tratamiento con etanol, los ciegos de ambos grupos fueron colectados, sometidos a preparación de membranas que después de coloreados por el método de Giemsa. Fue posible evaluar la densidad neuronal en un área de 13,44 mm2, y medir el área del cuerpo celular de 300 neuronas por grupo. Los ratones alcohólicos presentaron un incremento en el número de neuronas pequeñas y una disminución de las neuronas medianas y grandes. La densidad neuronal verificada en los ratones alcohólicos fue significativamente reducida, en relación a los ratones controles. Esa reducción dejó de ser estadísticamente significativa cuando fue proyectada la densidad de las neuronas para el área del ciego, ya que la observación macroscópica mostraba que los ratones alcohólicos presentaban un ciego dilatado. El alcoholismo indujo una reducción significativa en el peso corporal final de los ratones del grupo A, provocó dilatación del ciego de los ratones, llevando a una gran dispersión neuronal. La dilatación del ciego de los ratones alcohólicos está probablemente asociada a alteraciones funcionales de las neuronas mientéricas que repercuten en el tono de la musculatura lisa intestinal.

Analysis of myenteric neurons of the cecum of diabetic rats after supplementation with ascorbic acid / Análisis de neuronas mientéricas del ciego de ratas diabéticas después de suplementación con ácido ascórbico

The objective of this work was to investigate the neuroprotective action of the ascorbic acid over the myenteric neurons in the cecum of Wistar rats, four months after induction of the diabetes mellitus experimental with streptozotocin. Three groups with five rats each were used: C- controls, D- diabetic, DA- diabetic treated with ascorbic acid. For evidentiation of the myenteric neurons was carried out to Giemsa's technique. Were evaluated the areas of cell bodies of 500 neurons in each group studied. The quantitative analysis was carried out in an area of 16.6 mm2 in each cecum studied. In the animals diabetic observed elevation of the glycemia and glycated hemoglobin. The supplementation with ascorbic acid was effective under the myenteric neurons of the cecum of diabetics rays, since was presented the effect neuroprotective and neurotrofic.

El objetivo de este trabajo fue verificar el efecto neuroprotector del ácido ascórbico sobre las neuronas mientéricas en el ciego de Rattus Wistar, cuatro meses después de la inducción de diabetes mellitus experimental con estreptozotocina. Utilizamos tres grupos de animales: C- control, D- diabético, DA- diabético tratado con ácido ascórbico. Para la observación de las neuronas mientéricas fue llevado a cabo la técnica de Giemsa. Fueron evaluadas las áreas del soma de 500 neuronas, en cada grupo estudiado. El análisis cuantitativo fue llevado a cabo, en cada ciego, en un área de 16,6 mm². En los animales diabéticos, se observó la elevación de la glicemia y de la hemoglobina glicosilada. La suplementación con ácido ascórbico fue efectiva en las neuronas mientéricas del ciego de animales diabéticos, ya que se produjeron los efectos neuroprotetor y neurotrófico.

Density and morphometry of overall myenteric neurons population from the duodenum of diabetic rats (Rattus norvegicus) under ascorbic acid supplementation / Densidad y morfometría de la población total de neuronas mientéricas del duodeno de ratones diabéticos (Rattus norvegicus) tratados con ácido ascórbico

The objective of this work was to evaluate the overall myenteric neurons population from the duodenum of adult streptozotocin-induced diabetic rats supplemented with ascorbic acid (AA), a potent antioxidant. Fifteen 90-day-old rats were divided in groups: control (C), diabetic (D), diabetic treated with ascorbic acid (DA). After 120 days of experimental period duodenums were resected and processed as whole-mount preparations according to Giemsa's technique, which allowed us to evaluate neuronal density in an area of 8.96 mm² and measure the area of 500 neuronal cell bodies per group. It was observed a 32.55% reduction in neuronal density of group D when compared to group C (p<0.05). The density of spared neurons in group DA, in relation to group D, was not statistically different in this experimental model. No significant differences were found in neuronal areas when groups C and D or group D and DA were compared (p>0.05). These results lead us to conclude that the density of overall myenteric neurons population from the duodenum was reduced in diabetic rats (D), when compared to its control (C); and that diabetic rats supplemented with AA (DA) did not have their neuronal density preserved when compared to diabetic animals (D).

El objetivo de este trabajo fue evaluar la población total de neuronas mientéricas del duodeno de ratones adultos inducidos a diabetes por estreptozotocina, suplementados con ácido ascórbico (AA), un poderoso antioxidante. Quince ratones con 90 días de edad fueron divididos en los grupos: control (C), diabético (D) y diabético tratados con ácido ascórbico (DA). Después de 120 días de tratamiento con AA, los duodenos fueron resecados y procesados con el método de Giemsa, el cual permitió evaluar la densidad neuronal, en un área de 8,96 mm², y medir el área del soma de 500 neuronas por grupo. Se observó una reducción de 32,55% de la densidad neuronal del grupo D con respecto grupo C (p<0,05). La densidad de las neuronas observada en el grupo DA, en relación con el grupo D, no fue estadísticamente significativa en este modelo experimental. No fueron encontradas diferencias significativas en las áreas de neuronas, cuando los grupos C y D o el grupo D y DA (p>0,05) fueron comparados. Nuestros resultados permitieron concluir que la densidad de la población total de las neuronas mioentéricas del duodeno estuvo reducida en los ratones diabéticos comparados con los controles, mientras que, los ratones diabéticos suplementados con AA no mantuvieron su densidad neuronal cuando fueron comparados con los animales del grupo diabético.

Effects of the ascorbic acid supplementation on NADH-diaphorase myenteric neurons in the duodenum of diabetic rats.

We assessed the ascorbic acid (AA) supplementation on the myenteric neurons in the duodenum of rats. Fifteen rats with 90 days of age were divided into three groups: control (C), diabetics (D) and ascorbic acid treated diabetics (DA). After 120 days of daily treatment with AA, the duodenum was submitted to the NADH-diaphorase (NADH-d) histochemical technique, which allowed us to evaluate the neuronal density in an area of 8.96 mm2 for each duodenum, and also to measure the cellular profile area of 500 neurons per group. The supplementation promoted an increase on AA levels. The neuronal density (p < 0.05) was higher in the group DA when compared to group D. There were no significant differences in the neuronal areas, when we compared groups C (204 +/- 16.5) and D (146.3 +/- 35.84) to groups D and DA (184.5 +/- 5.6) (p > 0.05). The AA-supplementation avoided the density reduction of the NADHd myenteric neurons in the duodenum of diabetic rats.

Morphological and quantitative analyses of the NADH-d myenteric neurons of the jejunum of rats submitted to experimental chronic alcoholism / Análisis Morfológico y Cuantitativo de de Neuronas Mientéricas NADH-d del Yeyuno de Ratas Sometidas a Alcoholismo Crónico Experimental

El objetivo del trabajo fue verificar, durante un período experimental de 210 días, los posibles efectos del etanol sobre la morfología y densidad de las neuronas mientéricas NADH-diaforasa, en el yeyuno de ratas alcohólicas. Utilizamos 10 animales (Rattus norvegicus) separadas en dos grupos: el control (n=5) que recibió una dieta proteica normal (22 por ciento) y agua ad libitum; el otro, alcohólico, que recibió dieta proteica normal NUVILAB (22 por ciento) y brandy de azúcar de caña diluido a 30 Gay Lussac (30 v/v). El segmento de yeyuno fue obtenido y sometido a técnicas histoquímicas para teñir las células nerviosas. La observación a través de microscopía de luz mostró que no hubo diferencias morfológicas de importancia entre las neuronas del grupo control y el sometido a alcoholismo. El recuento neuronal en el grupo control, llevado a cabo en 40 campos microscópicos (8,96 µm2), de las regiones mesentérica y antimesentérica, determinó 1.131 y 693 neuronas, respectivamente, mientras que en el grupo alcohólico se encontraron 1.229 y 860 neuronas, respectivamente. El incremento significativo en el número de neuronas en la región mesentérica, en el grupo de las ratas alcohólicas, es debido a un crecimiento físico menor de esos animales comparados con el grupo control. El etanol causó malnutrición y consecuentemente, las ratas alcohólicas mostraron una densidad neuronal más amplia debido a una dispersión menor.

The objective of our work was to verify, during an experimental period of 210 days, the possible effects of the ethanol on the morphology and density of the NADH-diaforase myenteric neurons in the jejunum of alcoholic rats. We used 10 animals (Rattus norvegicus) separated in 2 groups: the controls (n=5), that received a normal proteic diet (22%) and water ad libitum; the alcoholic, that received NUVILAB normal proteic chow (22%) and sugar cane brandy diluted at 30 Gay Lussac (30 v/v). The jejunum segment was collected and submitted to the histochemical technique to stain the nervous cells and, then, to the elaboration of membrane whole mounts. The observation through light microscopy showed that there are no expressive morphologic differences between the ganglia of neurons of the control and alcoholic rats. The counting of neurons, carried out in 40 microscopic fields (8.96µm2) in the control group, at the mesenteric and antimesenteric regions, found 1,131 and 693 neurons respectively, while, the alcoholic group found 1,229 and 860 neurons. The significant increase in the number of neurons in the mesenteric region, in the alcoholic rats, is due to the smaller physical growth of those animals when compared to the controls. The ethanol caused malnutrition and consequently the alcoholic rats showed a larger neuronal density due to its smaller dispersion.

Profile of the cell body of nadh-diaphorase positive myenteric neurons from the stomach of rats (Rattus norvegicus) subjected to chronic alcoholism

O objetivo do trabalho foi comparar o perfil dos corpos celulares dos neurônios mioentéricos NADH-diaforase positivos do estômago de ratos. Utilizou-se 10 animais (Rattus norvegicus), provenientes dos grupos: a) controle (n=5), que durante 210 dias receberam, ad libitum, dieta com teor protéico normal (22 por cento) e água; e b) experimental (n=5), que durante 210 dias receberam, ad libitum, ração com teor protéico normal (22 por cento) e aguardente-de-cana diluída a 30 Gay Lussac (30º v/v). Os estômagos coletados foram submetidos à técnica de evidenciação neuronal. A mensuração do perfil dos corpos celulares dos neurônios (n=1.000) foi através de um Sistema Computadorizado de Análise de Imagem. O perfil dos corpos celulares dos neurônios do grupo controle ficou entre 60,16 a 638,64 m2. No grupo experimental variou de 40,84 a 599,15 µm2. Constatamos redução significante no tamanho do corpo celular, aumento de neurônios pequenos e diminuição de neurônios grandes

Density and morphometry of myenteric neurons of the ileum of rats subjected to chronic alcoholism

This work verified the changes caused by ethanol ingestion on the myenteric neurons of the ileum of rats subjected to chronic alcoholism. Twenty rats (Rattus norvegicus) were divided in control (GC) and alcoholic (GA) groups during 210 days. The animals from group GA received sugar-cane brandy diluted to 30 GL and those from group GC received water. Both groups were fed with rodent chow ad libitum. The ileum was prepared as whole-mounts for neuronal staining according to Barbosa (1978) and Gabella (1979). The neurons were counted and the cell bodies were measured. The statistical analysis revealed a significant decrease in the number of large neurons in GA compared with GC, while there was a significant increase in the number of small neurons in GA. There was a smaller body growth in GA, thus causing a smaller spread of the nerve cells and a larger number of neurons evidenced by the techniques of Giemsa and NADH-diaphorase in the alcoholic animals than in the controls, and a decrease in the size of the cell bodies, evidenced by the increase in the proportion of small neurons and decrease in the proportion of large nerve cells.

Localização dos neurônios mientéricos no estômago aglandular e glandular de ratos (rattus norvegicus) / Localization of the myenteric neurons in the aglandular and glandular stomach rats (rattus norvegicus)

O sistema nervoso entérico tem dentre suas funções, o papel de controlar a motricidade intestinal. É formado por gânglios intercalados à uma rede de fibras nervosas dispostas entre os estratos musculares circular e longitudinal do tracto gastrointestinal. Dependendo da espécie animal e da parte do tracto gastrointestinal analisado, a literatura revela diferenças morfológicas em relação à localização do plexo mientérico. O objetivo deste trabalho foi analisar a localização dos neurônios mientéricos nas diferentes regiões do estômago de ratos: estômago aglandular e glandular. Do estômago de 5 ratos (Rattus norvegicus) foram retirados fragmentos e realizados cortes longitudinais de 7 e 10 µm de espessura e corados por Hematoxilina-Eosina, Verhoeff-Van Gieson, Azan, Tricrômico de Picrosírius-Hematoxilina. Os neurônios do plexo mientérico encontram-se geralmente agrupados em gânglios. Os gânglios são envoltos por tecido conjuntivo rico em fibras colágenas e escassas fibras elásticas. Estão contidos na túnica muscular, porém com diferentes localizações. Na região aglandular estão entre os estratos longitudinal e circular da túnica muscular; já, na região glandular são observados também no interior do estrato longitudinal. Verificamos, portanto, que o plexo mientérico não apresenta o mesmo padrão de localização dos seus gânglios quando se analisa as diferentes regiões que formam o estômago do rato.