RESUMO
This study aimed to determine whether the effects on the mouse liver caused by three periods of feeding a protein-free diet for 5 days followed by a normal complete diet for 5 days (3PFD-CD) are prevented by a constant methionine supply (3PFD+Met-CD). The expressions of carbonic anhydrase III (CAIII), fatty acid synthase (FAS), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and glutathione S-transferase P1 (GSTP1) were assessed by proteomics and reverse transcriptase-polymerase chain reactions. The liver redox status was examined by measuring the activities of superoxide dismutase (SOD) and catalase (CAT), as well as protein carbonylation. Because oxidative stress can result in apoptosis, the activity and content of caspase-3, as well as the x-linked inhibitor of the apoptosis protein (XIAP) and mitochondrial caspase-independent apoptosis inducing factor (AIF) contents were assessed. In addition, the liver histomorphology was examined. Compared to the controls fed a normal complete diet throughout, feeding with 3PFD-CD increased the FAS content, decreased the CAIII content, decreased both the SOD and CAT activities, and increased protein carbonylation. It also activated caspase-3, decreased the XIAP content, decreased the AIF content, increased the number of GSTP1-positive foci and caspase-3-positive cells, and caused fatty livers. Conversely, the changes were lessened to varying degrees in mice fed 3PFD+Met-CD. The present results indicate that a regular Met supply lessens the biochemical changes, damage, and caspase-dependent apoptosis provoked by recurrent dietary amino acid deprivation in the mouse liver.
Assuntos
Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Metionina/farmacologia , Deficiência de Proteína/enzimologia , Animais , Fator de Indução de Apoptose/metabolismo , Feminino , Glutationa S-Transferase pi/metabolismo , Gliceraldeído-3-Fosfato Desidrogenase (Fosforiladora)/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Deficiência de Proteína/patologia , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/metabolismoRESUMO
Diarrhea increases the effects of malnutrition. Accordingly, the effect of diarrhea on two types of malnutrition (protein deficiency and protein-calorie deficiency) was studied. The experiment included 42 young Sprague Dawley rats. The rats were distributed into three groups with 14 rats per group. During the first 16 of the experiment, the first group was fed a control diet ad libitum, the second received the same diet but with food intake reduced in 50% whereas the third group was offered a protein deficient diet. Thus, at the end of this period there were well-fed rats (control), as well as protein and protein-calorie malnourished rats. Then one half of the rats in each group were given lactose to produce diarrhea and all rats continued with their previously assigned diet and feeding regime during one more week. Therefore, during this period there were control rats, protein deficient rats and protein-calorie deficient rats with and without diarrhea. The results showed that diarrhea caused a substantial reduction in food intake and growth in the well-fed rats and also in the group fed the protein deficient diet. However, the protein-calorie deficient group did not reduce its intake nor its growth rate. As a result, diarrhea caused malnutrition in the control group and increased malnutrition in the protein deficient but it did not have an additional effect in the protein-calorie deficient rats. The apparent absorption of lipids and nitrogen measured in these rats showed that the absorption reduction caused by diarrhea was more pronounced in the protein deficient group. This group also had the lowest activities of intestinal disaccharidases. These results showed that diarrhea had a more detrimental effect in protein deficient than in protein-calorie deficient rats.
Assuntos
Diarreia/metabolismo , Absorção Intestinal/fisiologia , Intestino Delgado/metabolismo , Deficiência de Proteína/metabolismo , Animais , Metabolismo dos Carboidratos , Diarreia/etiologia , Diarreia/fisiopatologia , Dissacaridases/metabolismo , Modelos Animais de Doenças , Gorduras/metabolismo , Alimentos , Nitrogênio/metabolismo , Deficiência de Proteína/enzimologia , Deficiência de Proteína/fisiopatologia , Desnutrição Proteico-Calórica/enzimologia , Desnutrição Proteico-Calórica/metabolismo , Desnutrição Proteico-Calórica/fisiopatologia , Ratos , Ratos Sprague-DawleyRESUMO
La diarrea magnifica los efectos de la desnutrición. En consecuencia, aquí se estudió el efecto de la diarrea sobre dos tipos de desnutrición (proteica y proteico-calórica). El experimento incluyó 42 ratas jóvenes de la cepa Sprague Dawley que se distribuyeron en tres grupos (14 ratas/grupo). Durante los primeros 16 días del experimento, el primer grupo recibió una dieta control ad-libitum, el segundo recibió la misma dieta pero su consumo se redujo en un 50% y el tercer grupo recibió una dieta deficiente en proteína. Al final de este período había ratas bien nutridas (controles) y con desnutrición proteica y calórico-proteica. Luego, a la mitad de estas ratas en cada grupo, se les produjo diarrea con lactosa y todas las ratas continuaron con su dieta y el régimen de alimentación preasignado durante una semana. Así, durante este período había ratas controles así como con deficiencia proteica o calórico-proteica que tenían diarrea y grupos idénticos que no tenían diarrea. Los resultados mostraron que la diarrea causó una disminución del consumo y del crecimiento en las ratas del grupo control y deficiente en proteína. Sin embargo, el grupo con deficiencia calórico-proteica no redujo su consumo ni disminuyó su crecimiento en respuesta a la diarrea. La consecuencia de esto fue que la diarrea produjo desnutrición en el grupo control y aumentó la desnutrición en el grupo deficiente en proteína, pero no tuvo un efecto adicional en el grupo con deficiencia calórico-proteica. Además, la reducción en la absorción aparente del nitrógeno y de la grasa asociada con la diarrea, fue mayor en las ratas deficientes en proteína. Este grupo también presentó las actividades más bajas de disacaridasas intestinales. Esto resultados muestran que la diarrea tiene un efecto negativo mayor en ratas con deficiencia proteica que con deficiencia calórico-proteica.
Diarrhea increases the effects of malnutrition. Accordingly, the effect of diarrhea on two types of malnutrition (protein deficiency and protein-calorie deficiency) was studied. The experiment included 42 young Sprague Dawley rats. The rats were distributed into three groups with 14 rats per group. During the first 16 of the experiment, the first group was fed a control diet ad libitum, the second received the same diet but with food intake reduced in 50% whereas the third group was offered a protein deficient diet. Thus, at the end of this period there were well-fed rats (control), as well as protein and protein-calorie malnourished rats. Then one half of the rats in each group were given lactose to produce diarrhea and all rats continued with their previously assigned diet and feeding regime during one more week. Therefore, during this period there were control rats, protein deficient rats and protein-calorie deficient rats with and without diarrhea. The results showed that diarrhea caused a substantial reduction in food intake and growth in the well-fed rats and also in the group fed the protein deficient diet. However, the protein-calorie deficient group did not reduce its intake nor its growth rate. As a result, diarrhea caused malnutrition in the control group and increased malnutrition in the protein deficient but it did not have an additional effect in the protein-calorie deficient rats. The apparent absorption of lipids and nitrogen measured in these rats showed that the absorption reduction caused by diarrhea was more pronounced in the protein deficient group. This group also had the lowest activities of intestinal disaccharidases. These results showed that diarrhea had a more detrimental effect in protein deficient than in protein-calorie deficient rats.
Assuntos
Animais , Ratos , Absorção Intestinal/fisiologia , Intestino Delgado/metabolismo , Deficiência de Proteína/metabolismo , Metabolismo dos Carboidratos , Diarreia , Modelos Animais de Doenças , Dissacaridases/metabolismo , Gorduras/metabolismo , Nitrogênio/metabolismo , Deficiência de Proteína/enzimologia , Deficiência de Proteína/fisiopatologia , Desnutrição Proteico-Calórica/enzimologia , Desnutrição Proteico-Calórica/metabolismo , Desnutrição Proteico-Calórica/fisiopatologia , Ratos Sprague-DawleyRESUMO
In humans and other animals, it has been shown that protein malnutrition during the prenatal period leads to permanent changes, which in adulthood may cause chronic diseases. Molecules involved in the control of energy metabolism could be targets to alterations caused by nutritional status. Some hypothalamic nuclei as the paraventricular (PVN), ventro-medial and arcuate are related to energy metabolism regulation. Orexigenic and anorexigenic molecules are involved in this regulation. Some studies have showed that these nuclei present nitric oxide synthase (NOS) and that it is increased in obese rats. Recently it had been shown that rats malnourished during the lactation period presented metabolic alterations that persist in adulthood. The aim of this work was to study the expression of NOS in hypothalamic nuclei of rats submitted to malnutrition during the early lactation period. Rats from post-natal day (P10) to P90 were used. Control dams were fed with regular chow pellets and diet dams were fed with protein-free chow pellets during the first 10 days of lactation. NADPH-diaphorase or immunostaining techniques were used to access NOS expression in hypothalamic nuclei. Our results show a delay in NOS expression in the PVN and VMH of malnourished rats. It may affect the development of the hypothalamic circuitry, leading to a metabolic imprinting.
Assuntos
Hipotálamo/enzimologia , Hipotálamo/crescimento & desenvolvimento , Lactação , Óxido Nítrico Sintase/análise , Deficiência de Proteína/enzimologia , Envelhecimento , Animais , Hipotálamo Médio/enzimologia , Imuno-Histoquímica , Masculino , NADPH Desidrogenase/análise , Núcleo Hipotalâmico Paraventricular/enzimologia , Ratos , Núcleo Supraóptico/enzimologiaRESUMO
Protein malnutrition leads to functional impairment in several organs, which is not fully restored with nutritional recovery. Little is known about the role of oxidative stress in the genesis of these alterations. This study was designed to assess the sensitivity of blood oxidative stress biomarkers to a dietary protein restriction. Male Wistar rats were divided into two groups, according to the diet fed from weaning (21 days) to 60 day old: normal protein (17% protein) and low protein (6% protein). Serum protein, albumin, free fatty acid and liver glycogen and lipids were evaluated to assess the nutritional status. Blood glutathione reductase (GR) and catalase (CAT) activities, plasma total sulfhydryl groups concentration (TSG) as well as plasma thiobarbituric acid reactive substances (TBARs) and reactive carbonyl derivatives (RCD) were measured as biomarkers of the antioxidant system and oxidative damage, respectively. The glucose metabolism in soleus muscle was also evaluated as an index of stress severity imposed to muscular mass by protein malnutrition. No difference was observed in muscle glucose metabolism or plasma RCD concentration between both groups. However, our results showed that the low protein group had higher plasma TBARs (62%) concentration and lower TSG (44%) concentration than control group, indicating increased reactive oxygen species production in low protein group. The enhancement of erythrocyte GR (29%) and CAT (28%) activities in this group also suggest an adaptation to the stress generated by the protein deficiency. Taken together, the results presented here show that the biomarkers used were able to reflect the oxidative stress level induced by this specific protein deficient diet.
Assuntos
Glucose/metabolismo , Estresse Oxidativo/fisiologia , Deficiência de Proteína/sangue , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Adaptação Fisiológica/fisiologia , Animais , Biomarcadores , Catalase/sangue , Glutationa Redutase/sangue , Masculino , Estado Nutricional , Deficiência de Proteína/enzimologia , Ratos , Ratos Wistar , Compostos de Sulfidrila/sangueRESUMO
Se analiza las consecuencias provocadas por desequilibrios nutricionales sobre el contenido de DNA y la actividad de las enzimas ADA y PNP en timo de ratas en crecimientos. Ratas Wistar al destete fueron alimentadas con: 1- dieta libre de proteínas hasta asemejar cuadros de malnutrición proteica leve, moderada y severa; 2- dieta conteniendo harina de maíz en baja concentración (6,5 por ciento). La subnutrición durante la lactancia se obtuvo duplicando la camada (12-14 crías por madre). Como controles se utilizaron ratas bien nutridas de igual edad, que desde el destete recibieron dieta stock. Al finalizar la experiencia, se les extrajo el timo (Pt)(mg). Se determinó DNA (mg/órgano), el número de núcleos, el tamaño celular- Pt(mg)/No. de Núcleos- y la actividad de las enzimas ADA y PNP (umol de ácido úrico x 10 - 1/P)(P=Pt(mg) /P corporal 0.75). Los resultados muestran que tanto la subnutrición durante la lactancia, como la malnutrición proteica al destete y la administración de dieta de baja calidad, afectan la proliferación celular en el timo, Sólo la carencia de proteína o su baja calidad, aumenta la actividad de ADA y PNP
Assuntos
Animais , Ratos , Deficiência de Proteína/enzimologia , Desnutrição Proteico-Calórica/enzimologia , Adenosina Desaminase/deficiência , Adenosina Desaminase/farmacologia , Dieta com Restrição de Proteínas/efeitos adversos , Purina-Núcleosídeo Fosforilase/deficiência , Purina-Núcleosídeo Fosforilase/farmacologia , Ratos Wistar/crescimento & desenvolvimento , Análise de Sequência de DNA , Timo/enzimologiaRESUMO
The experiment was performed in order to evaluate the beta-glucuronidase activity in gastric juice and gastric mucosa of rats submitted to protein-free diet. A group of 36 young adult male wistar rats was fed a protein-free diet ad libitum for five weeks; a second group of 36 wistar rats ingested a purified isocaloric 12,5% casein diet for the same period. The concentration of proteins in plasma, gastric juice and gastric glandular mucosa and the beta-glucuronidase activity in the gastric juice and gastric glandular mucosa were determined. Protein deficient rats had lower plasma protein concentration and also a lower protein concentration in gastric juice and gastric mucosa. In these animals there was no significant change of beta-glucuronidase activity in the gastric juice, but there was a significant increase of the specific enzimatic activity in the gastric mucosa. The results suggest that protein restriction in young adult rats affects the gastric mucosa. The increase of the specific beta-glucuronidase activity might be due to heightened local catabolism or to a comparatively more severe protein depletion.
Assuntos
Suco Gástrico/enzimologia , Mucosa Gástrica/enzimologia , Glucuronidase/metabolismo , Deficiência de Proteína/enzimologia , Animais , Proteínas Sanguíneas/análise , Peso Corporal , Concentração de Íons de Hidrogênio , Masculino , RatosAssuntos
Fosfatase Alcalina/metabolismo , Dipeptidases/metabolismo , Jejuno/enzimologia , Deficiência de Proteína/enzimologia , Proteínas Sanguíneas/metabolismo , Colômbia , Estabilidade de Medicamentos , Humanos , Mucosa Intestinal/enzimologia , Jejuno/patologia , Masculino , Relação Estrutura-AtividadeAssuntos
Aminoácidos/metabolismo , Fígado/enzimologia , Liases/metabolismo , Distúrbios Nutricionais/enzimologia , Adaptação Fisiológica , Peso Corporal , Dietoterapia , Edema/enzimologia , Feminino , Hepatomegalia/enzimologia , Humanos , Lactente , Jamaica , Masculino , Deficiência de Proteína/enzimologiaRESUMO
Aminoacid-activating enzymes and argininosuccinase were measured in liver-biopsy samples from twenty-two malnourished and recovering Jamaican infants. Aminoacid-activating-enzyme levels were increased initially and fell on recovery; argininosuccinase activity was low initially and rose on recovery. It is suggested that these changes are part of the adaptation which takes place in aminoacid metabolism in the malnourished state. (AU)